Method of treatment and prevention of bacterial vaginosis

ABSTRACT

Described herein are methods of reducing the risk of recurrence of bacterial vaginosis by intravaginal administration of compositions comprising L-lactic acid, polymer thickener, and preservative.

CROSS-REFERENCE

This application is a continuation of U.S. application Ser. No.16/338,977, now U.S. Pat. No. 11,419,835, which is a 371 national stageentry of International Patent Application PCT/US2017/054943, filed Oct.3, 2017, which claims the benefit of U.S. Provisional Application No.62/404,061 filed Oct. 4, 2016, each of which is incorporated byreference herein in its entirety.

BACKGROUND OF THE DISCLOSURE

There is a need for developing compositions and methods for improvedtreatment and prevention of bacterial vaginosis (BV).

SUMMARY OF THE DISCLOSURE

Disclosed herein, in some embodiments, are methods of treating bacterialvaginosis (BV) comprising intravaginally administering a composition toa subject with a BV, wherein the composition comprises: (a) a polymerthickener; (b) L-lactic acid; and (c) a preservative, whereinadministering the composition ameliorates symptoms of BV in the subject.In some instances, the polymer thickener is selected from a groupconsisting of xanthan gum, alginic acid, hydroxypropyl cellulose,hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose,chitosan, polycarbophil, and carbopol. In some instances, thecomposition further comprises a humectant. In some instances, thecomposition further comprises at least one of a pharmaceuticallyacceptable carrier, water, and a buffer. In some instances, the buffercomprises citric acid and potassium bitartrate. In some instances, thecomposition is administered once or multiple times during a course oftreatment. In some instances, the course of treatment comprises about 1week to about 20 weeks. In some instances, the course of treatmentcomprises an initial course of treatment and a subsequent course oftreatment and wherein the composition is administered more frequentlyduring the initial course of treatment than during the subsequent courseof treatment. In some instances, the frequency of administration duringthe initial course of treatment is daily, less than once daily, everyother day, once a week, or once every 2 weeks. In some instances, thefrequency of administration during the subsequent course of treatment isless than once daily, every other day, once a week, once every 2 weeks,or once every 3 weeks. In some instances, the composition isadministered at a dosage from about 0.5 g to about 10 g, from about 3 gto about 5 g, about 3 g, about 4 g, or about 5 g. In some instances,administration of the composition lowers vaginal pH of the subject,thereby treating the BV. In some instances, administration of thecomposition restores a healthy vaginal microbiome in the subject,thereby treating the BV. In some instances, the subject has previouslybeen diagnosed as Amsel-positive. In some instances, upon ofadministration of the composition the subject is Amsel-negative, therebytreating the BV.

Disclosed herein, in some embodiments, is a method of reducing therecurrence rate of bacterial vaginosis (BV) comprising intravaginallyadministering a composition to a subject, wherein the compositioncomprises: (a) a polymer thickener; (b) L-lactic acid; and (c) apreservative, wherein administering the composition reduces recurrencerate of bacterial vaginosis in the subject. In some instances, thesubject is a subject with BV. In some instances, the subject with BV isasymptomatic In some instances, the subject is free of bacterialvaginosis (BV). In some instances, the polymer thickener is selectedfrom a group consisting of xanthan gum, alginic acid, hydroxypropylcellulose, hydroxypropyl methyl cellulose, sodium carboxymethylcellulose, chitosan, polycarbophil, and carbopol. In some instances, thecomposition further comprises a humectant. In some instances, thecomposition further comprises at least one of a pharmaceuticallyacceptable carrier, water, and a buffer. In some instances, the buffercomprises citric acid and potassium bitartrate. In some instances, thecomposition is administered once or multiple times during a course oftreatment. In some instances, the course of treatment comprises about 1week to about 20 weeks. In some instances, the course of treatmentcomprises an initial course of treatment and a subsequent course oftreatment and wherein the composition is administered more frequentlyduring the initial course of treatment than during the subsequent courseof treatment. In some instances, the frequency of administration duringthe initial course of treatment is daily, less than once daily, everyother day, once a week, or once every 2 weeks. In some instances, thefrequency of administration during the subsequent course of treatment isless than once daily, every other day, once a week, once every 2 weeks,or once every 3 weeks. In some instances, the composition isadministered at a dosage from about 0.5 g to about 10 g, from about 3 gto about 5 g, about 3 g, about 4 g, or about 5 g. In some instances,administration of the composition lowers vaginal pH of the subject,thereby reducing the recurrence rate of the BV. In some instances,administration of the composition lowers vaginal pH of the subject,thereby reducing the recurrence rate of the BV. In some instances,administration of the composition restores a healthy vaginal microbiomein the subject, thereby reducing the recurrence rate of the BV. In someinstances, the subject has previously been diagnosed as Amsel-positive.In some instances, upon of administration of the composition the subjectis Amsel-negative, thereby reducing the recurrence rate of the BV.

In one embodiment is provided a method of treating bacterial vaginosis(BV) comprising intravaginally administering a composition to a subjectwith BV, wherein the composition comprises: (a) a polymer thickener; (b)L-lactic acid; and (c) a preservative, wherein administering thecomposition ameliorates symptoms of BV in the subject. In anotherembodiment is provided a method of reducing the recurrence rate ofbacterial vaginosis (BV) comprising intravaginally administering acomposition to a subject with BV, wherein the composition comprises: (a)a polymer thickener; (b) L-lactic acid; and (c) a preservative, whereinadministering the composition reduces recurrence rate of bacterialvaginosis in the subject. Yet another embodiment provides a method ofpreventing the recurrence of bacterial vaginosis (BV) comprisingintravaginally administering a composition to a subject with BV, whereinthe composition comprises: (a) a polymer thickener; (b) L-lactic acid;and (c) a preservative, wherein administering the composition preventsrecurrence of bacterial vaginosis in the subject. In some embodiments,the preservative is selected from a group consisting of benzoic acid,sodium benzoate, methylparaben, ethylparaben, butyulparaben,propylparaben, benzyalkonium chloride, phenylmercuric nitate, andchlorhexidine. In some embodiments, the polymer thickener is selectedfrom a group consisting of xanthan gum, alginic acid, hydroxypropylcellulose, hydroxypropyl methyl cellulose, sodium carboxymethylcellulose, chitosan, polycarbophil, and carbopol. In some embodiments,the polymer thickener is a combination of xanthan gum and alginic acid.In some embodiments, the composition further comprises a humectant. Insome embodiments, the humectant is selected from a group consisting ofglycerol, polyethylene glycol, propylene glycol, sorbitol, andtiracetin. In some embodiments, the composition further comprises apharmaceutically acceptable carrier. In some embodiments, thecomposition further comprises water. In some embodiments, thecomposition further comprises a buffer. In some embodiments, the buffercomprises citric acid and potassium bitartrate.

In some embodiments, the composition is administered once during acourse of treatment. In some embodiments, the composition isadministered multiple times during a course of treatment. In someembodiments, the course of treatment comprises an initial course oftreatment and a subsequent course of treatment and wherein thecomposition is administered more frequently during the initial course oftreatment than during the subsequent course of treatment. In someembodiments, the initial course of treatment is between 1 week and 4weeks. In some embodiments, the initial course of treatment is about 2weeks to about 3 weeks. In some embodiments, the initial course oftreatment is about 1 week. In some embodiments, the subsequent course oftreatment is about 1 week to about 19 weeks. In some embodiments, thesubsequent course of treatment is about 2 weeks to about 18 weeks. Insome embodiments, the subsequent course of treatment is about 3 weeks toabout 17 weeks. In some embodiments, the subsequent course of treatmentis about 4 weeks to about 16 weeks. In some embodiments, the subsequentcourse of treatment is 5 weeks to about 15 weeks. In some embodiments,the subsequent course of treatment is about 6 weeks to about 14 weeks.In some embodiments, the subsequent course of treatment is about 7 weeksto about 13 weeks. In some embodiments, the subsequent course oftreatment is about 8 weeks to about 12 weeks. In some embodiments, thesubsequent course of treatment is about 9 weeks to about 10 weeks. Insome embodiments, the subsequent course of treatment is about 10 weeksto about 11 weeks. In some embodiments, the frequency of administrationduring the initial course of treatment is daily. In some embodiments,the frequency of administration during the initial course of treatmentis every other day. In some embodiments, the frequency of administrationduring the initial course of treatment is once a week. In someembodiments, the frequency of administration during the initial courseof treatment is once every 2 weeks. In some embodiments, the frequencyof administration during the subsequent course of treatment is everyother day. In some embodiments, the frequency of administration duringthe initial course of treatment is once every week. In some embodiments,the frequency of administration during the initial course of treatmentis once every 2 weeks. In some embodiments, the frequency ofadministration during the initial course of treatment is once every 3weeks. In some embodiments, the composition is administered at a dosagefrom about 0.5 g to about 10 g. In some embodiments, the composition isadministered at a dosage from about 3 g to about 5 g. In someembodiments, the composition is administered at a dosage selected fromabout 3 g, about 4 g, and about 5 g.

A further embodiment provides a method of prognosis for risk ofrecurrence of bacterial vaginosis (BV) in a subject with BV, the methodcomprising (a) testing vaginal pH of the subject; (b) intravaginallyadministering a composition as defined in any one of the aforementionedembodiments to the subject, (c) re-testing vaginal pH of the subject;and (d) prognosing risk of recurrence of BV based on comparing theresults of the vaginal pH tests in step (a) and (c). Another embodimentprovides a method of prognosis for risk of recurrence of bacterialvaginosis (BV) in a subject with BV, the method comprising (a) analyzingvaginal microbiome of the subject; (b) intravaginally administering acomposition as defined in any of the aforementioned embodiments to thesubject, (c) re-analyzing vaginal microbiome of the subject; and (d)prognosing risk of recurrence of BV based on comparing the results ofthe vaginal microbiome analyses in step (a) and (c). Yet anotherembodiment provides a method of prognosis for risk of recurrence ofbacterial vaginosis (BV) in a subject, the method comprising (a)assessing BV in a subject using Amsel criteria; (b) intravaginallyadministering a composition as defined in any one of the aforementionedembodiments to the subject, (c) re-assessing BV in the subject usingAmsel criteria; and (d) prognosing risk of recurrence of BV based oncomparing the assessments in step (a) and (c). In some embodiments, thecomposition is administered once during a course of treatment. In someembodiments, the composition is administered multiple times during acourse of treatment. In some embodiments, the course of treatmentcomprises an initial course of treatment and a subsequent course oftreatment and wherein the composition is administered more frequentlyduring the initial course of treatment than during the subsequent courseof treatment. In some embodiments, the initial course of treatment isbetween 1 week and 4 weeks. In some embodiments, the initial course oftreatment is about 2 weeks to about 3 weeks. In some embodiments, theinitial course of treatment is about 1 week. In some embodiments, thesubsequent course of treatment is about 1 week to about 19 weeks. Insome embodiments, the subsequent course of treatment is about 2 weeks toabout 18 weeks. In some embodiments, the subsequent course of treatmentis about 3 weeks to about 17 weeks. In some embodiments, the subsequentcourse of treatment is about 4 weeks to about 16 weeks. In someembodiments, the subsequent course of treatment is 5 weeks to about 15weeks. In some embodiments, the subsequent course of treatment is about6 weeks to about 14 weeks. In some embodiments, the subsequent course oftreatment is about 7 weeks to about 13 weeks. In some embodiments, thesubsequent course of treatment is about 8 weeks to about 12 weeks. Insome embodiments, the subsequent course of treatment is about 9 weeks toabout 10 weeks. In some embodiments, the subsequent course of treatmentis about 10 weeks to about 11 weeks. In some embodiments, the frequencyof administration during the initial course of treatment is daily. Insome embodiments, the frequency of administration during the initialcourse of treatment is every other day. In some embodiments, thefrequency of administration during the initial course of treatment isonce a week. In some embodiments, the frequency of administration duringthe initial course of treatment is once every 2 weeks. In someembodiments, the frequency of administration during the subsequentcourse of treatment is every other day. In some embodiments, thefrequency of administration during the initial course of treatment isonce every week. In some embodiments, the frequency of administrationduring the initial course of treatment is once every 2 weeks. In someembodiments, the frequency of administration during the initial courseof treatment is once every 3 weeks. In some embodiments, the compositionis administered at a dosage from about 0.5 g to about 10 g. In someembodiments, the composition is administered at a dosage from about 3 gto about 5 g. In some embodiments, the composition is administered at adosage selected from about 3 g, about 4 g, and about 5 g. In someembodiments, the lowering of vaginal pH between steps (a) and (c) isassociated with good prognosis for reduced risk of recurrence of BV. Insome embodiments, the restoration of a healthy vaginal microbiomebetween steps (a) and (c) is associated with good prognosis for reducedrisk of recurrence of BV. In some embodiments, the assessment result isAmsel-positive or Amsel-negative in the steps (a) and (c). In someembodiments, the change in assessment result from Amsel-positive toAmsel-negative between steps (a) and (c) is associated with goodprognosis for reduced risk of recurrence of BV.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 illustrates an exemplary study design for a phase 1 randomizedplacebo controlled pilot study to determine the effect and duration ofan intravaginal composition, as described in the present disclosure, onvaginal pH.

FIG. 2 illustrates an exemplary study design for a phase 1 randomizedplacebo controlled pilot study to determine the effect and duration ofan intravaginal composition, as described in the present disclosure, onvaginal pH.

FIG. 3 is a graph illustrating mean vaginal pH at each assessment timepoint.

FIG. 4 is a graph illustrating mean change from baseline in vaginal pHat each assessment time point.

FIG. 5 is a graph illustrating percentage of subjects with positiveoverall Amsel results—all reporting subjects at each assessmenttimepoint.

DETAILED DESCRIPTION OF THE DISCLOSURE

While preferred embodiments of the present disclosure have been shownand described herein, it will be obvious to those skilled in the artthat such embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the present disclosure. It should beunderstood that various alternatives to the embodiments of the presentdisclosure described herein may be employed. It is intended that thefollowing claims define the scope of the present disclosure and thatmethods and structures within the scope of these claims and theirequivalents be covered thereby.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of skill in the artto which the present disclosure described herein belong. Allpublications, patents, and patent applications mentioned in thisspecification are hereby incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.All documents, or portions of documents, cited in the applicationincluding, without limitation, patents, patent applications, articles,books, manuals, and treatises are hereby expressly incorporated byreference in their entirety for any purpose.

The terminology used herein is for the purpose of describing particularcases only and is not intended to be limiting. As used herein, thesingular forms “a”, “an” and “the” include the plural forms as well,unless the context clearly indicates otherwise. Furthermore, to theextent that the terms “contains,” “containing,” “including”, “includes,”“having,” “has”, “with”, or variants thereof are used in either thedetailed description and/or the claims, such terms are intended to beinclusive in a manner similar to the term “comprising.”

The term “about” or “approximately” are meant to refer to values withinan acceptable error range for the particular value as determined by oneof ordinary skill in the art, which will depend in part on how the valueis measured or determined, e.g., the limitations of the measurementsystem. For example, “about” means, in some cases, within 1 or more than1 standard deviation, per the practice in the given value. Whereparticular values are described in the application and claims, unlessotherwise stated the term “about” should be assumed to mean anacceptable error range for the particular value, such as ±10% of thevalue modified by the term “about”.

The terms “treat,” “treating,” and “treatment” include alleviating orabrogating a disorder, disease, or condition; or one or more of thesymptoms associated with the disorder, disease, or condition; oralleviating or eradicating the cause(s) of the disorder, disease, orcondition itself. Desirable effects of treatment include, but are notlimited to, preventing occurrence or recurrence of disease, alleviationof symptoms, diminishing any direct or indirect pathologicalconsequences of the disease, preventing metastasis, decreasing the rateof disease progression, amelioration or palliation of the disease stateand remission or improved prognosis. The terms “prevent,” and“preventing,” include reducing the likelihood of occurrence of adisease, disorder, or condition.

The term “subject with bacterial vaginosis (By)” includes subjects whocurrently or previously had BV. In some instances, the subject currentlyhas BV but is asymptomatic. In some instances, the subject is currentlyfree of BV due to previously administered treatment for BV, such as, forexample, antibiotics.

Bacterial Vaginosis

Bacterial vaginosis (BV) is a common condition characterized bydysbiosis of the vaginal microbiota, such that the proportion of lacticacid-producing bacteria, primarily Lactobacillus sp., is greatly reducedand the number and diversity of facultative and strictly anaerobicbacteria, including species of Gardnerella, Prevotella and otherClostridialis taxa, and Gram-positive cocci, including Enterococcusfaecalis, is greatly increased. Studies have shown that the microbialflora of the vagina, i.e., its microbiome, is usually affected by the pHof the vaginal fluid, such that a lower (more acidic) pH favorsLactobacillus sp., whereas a higher (more basic) pH favors anaerobicorganisms that are associated with BV. BV is typically associated with avaginal pH>5, elevated vaginal levels of Interleukin-6 (IL-6), aproinflammatory cytokine, increased risk of Neisseria gonorrhoeae,Chlamydia trachomatis, Trichomonas vaginalis and herpes simplex virus(HSV) type 2 infections, preterm delivery and low birth weight infants.Treatment with antibiotics often fails to correct the microfloraimbalance, and recurrence of BV is common. Accordingly, there is needfor an improved treatment of BV to reduce the risk of recurrence. Thepresent disclosure provides a method of treating or preventing BV byadministration of a composition described herein, such that the risk ofrecurrence of BV is reduced following administration of the composition.

BV is clinically identified if at least three of the following fourAmsel criteria are met: 1) thin, gray/white discharge; 2) malodorous“fishy” discharge upon adding 10% potassium hydroxide to vaginalsecretions (also referred to as “whiff test”; 3) high vaginal pH (>4.5),and 4) identification of vaginal epithelial cells heavily coated withbacteria (i.e., “clue cells”). In the present disclosure, a subject isdefined as Amsel-positive if the subject meets at least three out of thefour Amsel criteria. Further, in the present disclosure a subject isdefined as Amsel-negative if the subject does not meet at least threeout of the four Amsel criteria.

Methods of Treatment

Provided herein in one embodiment is a method of treating or preventingBV comprising intravaginally administering a composition comprising: (a)a polymer thickener, (b) L-lactic acid, and (b) a preservative. In someembodiments, the method described herein comprises intravaginallyadministering to a subject a composition comprising (a) a polymerthickener; (b) L-lactic acid; and (c) a preservative, whereinadministering the composition reduces the risk of recurrence of BV inthe subject.

In some embodiments, the composition comprises about 1% to about 10%L-lactic acid. In some embodiments, the composition comprises about 1%to about 9% L-lactic acid. In some embodiments, the compositioncomprises about 1% to about 8% L-lactic acid. In some embodiments, thecomposition comprises about 1% to about 7% L-lactic acid. In someembodiments, the composition comprises about 1% to about 6% L-lacticacid. In some embodiments, the composition comprises about 1% to about5% L-lactic acid. In some embodiments, the composition comprises about1% to about 4% L-lactic acid. In some embodiments, the compositioncomprises about 1% to about 3% L-lactic acid. In some embodiments, thecomposition comprises about 1% to about 2% L-lactic acid. In someembodiments, the composition comprises about 1% to about 1.5% L-lacticacid. The percent unit of the components refers to % weight/weight (%w/w).

In some embodiments, the composition comprises (a) about 0.1% to about10% of a polymer thickener, (b) about 1% to about 10% L-lactic acid, and(c) about 0.1% to about 10% of a preservative. In certain embodiments,the composition comprises, (a) about 1% to about 7% of a polymerthickener, (b) about 1% to about 10% L-lactic acid, and (c) about 0.1%to about 2% of a preservative. In some embodiments, the compositioncomprises (a) about 1.5% to about 8% of a polymer thickener, (b) about1% to about 5% L-lactic acid, and (c) about 0.1% to about 0.5% of apreservative. In certain embodiments, the composition comprises (a)about 2% to about 5% of a polymer thickener, (b) about 1% to about 2.5%L-lactic acid, and (c) about 0.2% to about 0.4% of a preservative. Incertain embodiments, the composition comprises (a) about 2.5% to about4.5% of a polymer thickener, (b) about 1% to about 2% L-lactic acid, and(c) about 0.1% to about 0.2% of a preservative. In certain embodiments,the composition comprises (a) about 6.25% of a polymer thickener, (b)about 1% L-lactic acid, and (c) about 0.2% of a preservative. In certainembodiments, the composition comprises (a) about 5.25% of a polymerthickener, (b) about 1% L-lactic acid, and (c) about 0.2% of apreservative.

Non-limiting examples of polymer thickeners include alginic acid,chitosan, gellan gum, xanthan gum, poloxamer, hydroxypropyl cellulose,hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose,polycarbophil, carbopol and the like. In some embodiments, the polymerthickener is alginic acid and xanthan gum. In some embodiments, thepolymer thickener is alginic acid. Alginic acid is a generally linearglycouronan polymer containing a mixture of polysaccharide polymer ofbeta-D-mannuronate and alpha-L-guluronate residues. The molecular weightof the alginic acid, in some instances, is in the range of about 20,000to about 300,000 g/mole, about 20,000 to about 250,000 g/mole, about240,000 g/mole, about 25,000 to about 30,000 g/mole, about 30,000 toabout 35,000 g/mole, about 35,000 to about 40,000 g/mole, about 40,000to about 45,000 g/mole, about 45,000 to about 50,000 g/mole, about50,000 to about 55,000 g/mole, about 55,000 to about 60,000 g/mole,about 60,000 to about 65,000 g/mole, about 65,000 to about 70,000g/mole, about 70,000 to about 75,000 g/mole, about 75,000 to about80,000 g/mole, about 80,000 to about 85,000 g/mole, about 85,000 toabout 90,000 g/mole, about 95,000 to about 100,000 g/mole, about 125,000to about 150,000 g/mole, about 150,000 to about 200,000 g/mole, or about100,000 to about 200,000 g/mole. The average molecular weight of thealginic acid is, in some instances, in the range of about 20,000 toabout 300,000 Da, about 20,000 to about 250,000 Da, about 240,000 Da,about 25,000 to about 30,000 Da, about 30,000 to about 35,000 Da, about35,000 to about 40,000 Da, about 40,000 to about 45,000 Da, about 45,000to about 50,000 Da, about 50,000 to about 55,000 Da, about 55,000 toabout 60,000 Da, about 60,000 to about 65,000 Da, about 65,000 to about70,000 Da, about 70,000 to about 75,000 Da, about 75,000 to about 80,000Da, about 80,000 to about 85,000 Da, about 85,000 to about 90,000 Da,about 95,000 to about 100,000 Da, about 125,000 to about 150,000 Da,about 150,000 to about 200,000 Da, or about 100,000 to about 200,000 Da,about 180,000 Da, about 183,855 Da, about 185,000 Da, about 190,000 Da,or about 195,000 Da. In certain examples, the alginic acid comprisesguluronic acid (G) and mannuronic acid (M) residues or units. In someinstances, the alginic acid comprises about 65-75% G residues and about25-35% M residues. The ratio between the M and G residues, are in someexamples, from about 0.1 to 0.2, about 0.2 to about 0.3, about 0.3 toabout 0.4, about 0.4 to about 0.5, about 0.5 to about 0.6, about 0.6 toabout 0.7, about 0.7 to about 0.8, about 0.8 to about 1, about 0.2,about 0.21, about 0.22, about 0.23, about 0.24, about 0.25, about 0.26,about 0.27, about 0.28, about 0.29, about 0.30, about 0.31, about 0.32,about 0.33, about 0.34, about, 0.35, about 0.36, about 0.37, about 0.39,about 0.39, about 0.40, about 0.41, about 0.42, about 0.43, about 0.44,about 0.45, about 0.46, about 0.47, about 0.48, about 0.49, or about0.5.

Alginic acid forms insoluble alginates by interacting with monovalentand divalent cations (especially Na⁺, K⁺, and Ca⁺⁺) in seminal plasma.Alginates also swell in contact with water, thereby assisting inmaintaining the gel form of the composition within the vagina. It isalso contemplated that alginic acid or salts of alginic acid contributeto the acid buffering activity of the composition since they have a pHof about 1.5 to about 3.5 in an aqueous solution. Furthermore, in someembodiments the alginic acid acts as a bioadhesive and provides thecomposition with bioadhesive properties. Without being bound by anyparticular theory, it is believed that because of its high molecularweight, alginic acid is not absorbed by the body. Thus, its effect as apolymer thickener, a bioadhesive, and acid-buffering agent is maintainedso as long as the gel remains in the vagina. In some embodiments, thepolymer thickener is xanthan gum. In some embodiments, the polymerthickener is a combination of xanthan gum and alginic acid. In someembodiments, the alginic acid is PROTACID F120.

The composition, in some embodiments, further comprises a bioadhesivecompound. Non-limiting examples of bioadhesive compounds include,xanthan gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose,sodium carboxymethyl cellulose, chitosan, polycarbophil, carbopol, andthe like. In some embodiments, the bioadhesive compound is xanthan gum,a high molecular weight polysaccharide gum containing D-glucosyl,D-mannosyl, and D-glucosyluronic acid residues and varying proportionsof O-acetyl and pyruvic acid acetal. The primary structure of xanthangum is a cellulose backbone with trisaccharide side chains; therepeating unit is a pentasaccharide. Generally, the molecular weight isgreater than about 10⁶ g/mole.

In some embodiments, L-lactic acid is a buffering agent that acts tomaintain the pH of the vagina within its normal acidic range (i.e., a pHof less than about 5 and more preferably in the range of about 3.5 toabout 4.5). In some embodiments, the composition further comprises abuffer. Non-limiting examples of buffering agents include, but are notlimited to, citric acid, potassium acid tartrate, benzoic acid, alginicacid, sorbic acid, fumaric acid, ascorbic acid, stearic acid, oleicacid, tartaric acid, edetic acid ethylenediaminetetracetic acid, aceticacid, malic acid, and the like. In some instances, the acids are addedas free acids, hydrates, or pharmaceutically acceptable salts. In someinstances, the free acids are converted to the corresponding salts insitu (i.e., within the vagina). In some embodiments, several bufferingagents are included in the combination gel to provide increasedbuffering capacity. In some embodiments, alginic acid, functions as botha polymer thickener and a buffering agent. In some instances, alginicacid is not absorbed by the body, thereby having a longer lasting acidbuffering effect compared to the other buffering agents that areabsorbed by the body.

In some instances, the compositions of the present disclosure includeadditional excipients. In some instances, the additional excipientsinclude humectants. Suitable humectants include, but are not limited to,for example, glycerol, polyethylene glycols, propylene glycols,sorbitol, triacetin, and the like. In some embodiments, glycerol is usedas a humectant to prevent the formation of a dry film on the gel whenplaced within the vagina. In certain embodiments, glycerol also acts asa lubricant. Additionally, the combination gel, in some embodiments,also includes a preservative. Suitable preservatives include, but arenot limited to, for example, benzoic acid, sodium benzoate,methylparaben, ethylparaben, butylparaben, propylparaben, benzyalkoniumchloride, phenylmercuric nitrate, chlorhexidine, and the like. In someembodiments, the preservative is benzoic acid. In some embodiments, thecombination gel comprises benzoic acid, which is a preservative and alsocontributes to the buffering capacity of the combination gel.

In some embodiments, the composition further comprises one or morecosmetic ingredients. Such cosmetic ingredients include diluents,solvents, and adjuvants, for example; water, ethyl alcohol, isopropylalcohol, glycerin, glycerol propylene glycol, sorbitol, and other highmolecular weight alcohols. In addition, the composition, in certainembodiments, further comprises minor amounts of other additives, suchas, for example; stabilizers, surfactants, menthol, eucalyptus oil,other essential oils, fragrances, and the like. The selection andamounts of cosmetic ingredients, other additives, and blendingprocedures are carried out in accordance with techniques well-known inthe art.

In some embodiments, the pharmaceutical carrier is water. In someembodiments, the pharmaceutical carrier is aqueous based. Otherpharmaceutically acceptable carriers that are suitable for vaginaldelivery are well known and are used, in some examples, in place ofwater. One example of a suitable pharmaceutically acceptable carrier ispetrolatum, such as white petrolatum.

In one exemplary embodiment of the present disclosure, the compositionis further described as follows: the polymer thickener is alginic acidand xanthan gum; the preservative is benzoic acid; the humectant isglycerol; citric acid, potassium bitartrate, and L-lactic acid areacidic components; and water is the pharmaceutically acceptable carrier.In another embodiment, the composition contains xanthan gum, alginicacid, L-lactic acid, citric acid, benzoic acid, potassium bitartrate,glycerol, and water.

As discussed, L-lactic acid or other suitable buffering agents are usedto maintain the pH of the vagina within its normal acidic range (i.e., apH of less than about 5 and more preferably in the range of about 3.5 toabout 4.5). Lactic acid has two isomers, one is known as L-(+)-lacticacid or (S)-lactic acid and the other is D-(−)-lactic acid or (R)-lacticacid. Recent discovery has shown that the L form of lactic acid is morepotent in inactivating HIV than D or racemic lactic acid. It iscontemplated herein that the L form of lactic acid is more potent ininactivating HIV than D or racemic lactic acid. While the precisemechanism of how L-lactic acid inactivates HIV is unknown, thestereochemical dependent activity suggests that it acts on proteins.Accordingly, the composition used in the methods described hereincomprises L-lactic acid and provides an improved therapeutic effectcompared to a similar composition comprising D-lactic acid.

The composition of the present disclosure is in the form of a gel, asemi-solid, a cream, a lotion, and/or a liquid. In some embodiments, thecomposition is injected. In some embodiments, the composition isinstilled. In some embodiments, the composition is administered as atopical ointment applied to the lining of the vagina and/or cervixand/or rectum. In some instances, this is accomplished as a gel, cream,lotion, non-aqueous or aqueous solution used to flush the vaginal orrectal cavity, and/or a vaginal or rectal suppository. In otherembodiments, the composition is administered in a spray formulation.

In some embodiments, the compositions of the present disclosure aredelivered to the vagina of a mammal by any means known to those skilledin the art. Typical forms for delivery of the compositions include, forexample; creams, lotions, gels, foams, intravaginal devices such assponges and suppositories, and films. In some instances, the compositionis additionally used as personal care products, such as, for example,condom lubricants, and the like. In some instances, such lubricantscomprise commonly known ingredients such as, for example: humectants,e.g., glycerin, sorbitol, mannitol, glycols and glycol ethers; buffers,e.g., glucono-d-lactone; germicides or bactericides, e.g., chlorhexidinegluconate; preservatives, e.g., methylparaben; viscosifiers, e.g.,hydroxyethyl cellulose, etc.; other adjuvants, e.g., colors andfragrances; in addition to the compositions of the present disclosure.Those skilled in the art will recognize that the physical properties,e.g., viscosity, of such delivery forms may vary widely. For example, insome instances, the viscosity of a gel form of the composition of thepresent disclosure, e.g., 150,000 centipoise, is substantially higherthan the viscosity of lotion form of the composition of the presentdisclosure, e.g., 100 centipoise. Further details concerning thematerials, ingredients, proportions and procedures of such deliveryforms are selected in accordance with techniques known in the field.

In some embodiments, the composition described herein is intravaginallyadministered one or more times during a course of treatment. In someembodiments, the composition described herein is intravaginallyadministered once during a course of treatment. In some embodiments, thecourse of treatment is about 1 week to about 20 weeks. In someembodiments, the course of treatment is about 2 weeks to about 19 weeks.In some embodiments, the course of treatment is about 3 weeks to about18 weeks. In some embodiments, the course of treatment is about 4 weeksto about 17 weeks. In some embodiments, the course of treatment is about5 weeks to about 16 weeks. In some embodiments, the course of treatmentis about 6 weeks to about 15 weeks. In some embodiments, the course oftreatment is about 7 weeks to about 14 weeks. In some embodiments, thecourse of treatment is about 8 weeks to about 13 weeks. In someembodiments, the course of treatment is about 9 weeks to about 12 weeks.In some embodiments, the course of treatment is about 10 weeks to about11 weeks. In certain embodiments, the course of treatment is about 3weeks to about 16 weeks. In some embodiments, the course of treatment isabout 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about10 weeks about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks,about 15 weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23 weeks,about 24 weeks, about 7 months, about 8 months, about 9 months, about 10months, about 11 months, about 12 months, about 13 months, or from about6 months to about 24 months.

In some embodiments, the composition is intravaginally administeredduring the course of treatment daily, every other day, once a week, onceevery 2 weeks, once every 3 weeks, once every 4 weeks, once every 5weeks, once every 6 weeks, once every 7 weeks, once every 8 weeks, onceevery 9 weeks, once every 10 weeks, once every 11 weeks, once every 12weeks, once every 13 weeks, once every 14 weeks, once every 15 weeks,once every 16 weeks, once every 17 weeks, once every 18 weeks, onceevery 19 weeks, or once every 20 weeks.

In some embodiments, the course of treatment comprises an initial courseof treatment and a subsequent course of treatment. In some embodiments,the initial course of treatment is shorter than the subsequent course oftreatment. In some embodiments, the initial course of treatment is about1 week to about 19 weeks. In some embodiments, the initial course oftreatment is about 2 weeks to about 18 weeks. In some embodiments, theinitial course of treatment is about 3 weeks to about 17 weeks. In someembodiments, the initial course of treatment is about 4 weeks to about16 weeks. In some embodiments, the initial course of treatment is about5 weeks to about 15 weeks. In some embodiments, the initial course oftreatment is about 6 weeks to about 14 weeks. In some embodiments, theinitial course of treatment is about 7 weeks to about 13 weeks. In someembodiments, the initial course of treatment is about 8 weeks to about12 weeks. In some embodiments, the initial course of treatment is about9 weeks to about 11 weeks. In some embodiments, the initial course oftreatment is about 10 weeks to about 11 weeks. In some embodiments, theinitial course of treatment is about 1 week to about 4 weeks. In someembodiments, the initial course of treatment is about 2 weeks to about 3weeks. In some embodiments, the initial course of treatment is about 1week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks,about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19 weeks,about 20 weeks, about 21 weeks, about 22 weeks, about 23 weeks, about 24weeks, about 7 months, about 8 months, about 9 months, about 10 months,about 11 months, about 12 months, about 13 months, or from about 6months to about 24 months. In some embodiments, the subsequent course oftreatment is about 1 week to about 19 weeks. In some embodiments, thesubsequent course of treatment is about 2 week to about 18 weeks. Insome embodiments, the subsequent course of treatment is about 3 weeks toabout 17 weeks. In some embodiments, the subsequent course of treatmentis about 4 weeks to about 16 weeks. In some embodiments, the subsequentcourse of treatment is about 5 weeks to about 15 weeks. In someembodiments, the subsequent course of treatment is about 6 weeks toabout 14 weeks. In some embodiments, the subsequent course of treatmentis about 7 weeks to about 13 weeks. In some embodiments, the subsequentcourse of treatment is about 8 weeks to about 12 weeks. In someembodiments, the subsequent course of treatment is about 9 weeks toabout 10 weeks. In some embodiments, the subsequent course of treatmentis about 10 weeks to about 11 weeks. In some embodiments, the subsequentcourse of treatment is about 1 week to about 4 weeks. In certainexamples, the subsequent course of treatment is about 1 week, about 2weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks,about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks,about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, about 7months, about 8 months, about 9 months, about 10 months, about 11months, about 12 months, about 13 months, or from about 6 months toabout 24 months.

In embodiments wherein the course of treatment comprises an initial anda subsequent course of treatment, the frequency of intravaginaladministration of the composition is different between the initial andthe subsequent course of treatment. In some embodiments, the compositionis intravaginally administered more frequently during the initial courseof treatment than during the subsequent course of treatment. In someembodiments, the composition is intravaginally administered lessfrequently during the initial course of treatment than during thesubsequent course of treatment.

In some embodiments, the composition is intravaginally administeredduring the initial course of treatment daily, every other day, once aweek, once every 2 weeks, once every 3 weeks, once every 4 weeks, onceevery 5 weeks, once every 6 weeks, once every 7 weeks, once every 8weeks, once every 9 weeks, once every 10 weeks, once every 11 weeks,once every 12 weeks, once every 13 weeks, once every 14 weeks, onceevery 15 weeks, once every 16 weeks, once every 17 weeks, once every 18weeks, or once every 19 weeks. In some embodiments, the frequency ofadministration during the initial course is once a day, once every 12hours, once every 6 hours, once every 4 hours, or once every 2 hours. Insome embodiments, the frequency of administration during the initialcourse is such that there is no more than 24 hours gap betweenadministration of two doses. In some embodiments, the frequency ofadministration during the initial course is such that there is no morethan 48 hours gap between administration of two doses. In someembodiments, the frequency of administration during the initial courseis such that there is no more than 72 hours gap between administrationof two doses. In some embodiments, the frequency of administrationduring the initial course is such that there is no more than 96 hoursgap between administration of two doses. In some embodiments, thefrequency of administration during the initial course is such that thereis no more than 120 hours gap between administration of two doses. Insome embodiments, the frequency of administration during the initialcourse is such that there is no more than 168 hours gap betweenadministration of two doses. In other embodiments, the frequency ofadministration during the initial course is such that there is more than24 hours gap between administration of two doses. In other embodiments,the frequency of administration during the initial course is such thatthere is more than 48 hours gap between administration of two doses. Inother embodiments, the frequency of administration during the initialcourse is such that there is more than 72 hours gap betweenadministration of two doses. In other embodiments, the frequency ofadministration during the initial course is such that there is more than96 hours gap between administration of two doses. In other embodiments,the frequency of administration during the initial course is such thatthere is more than 120 hours gap between administration of two doses. Inother embodiments, the frequency of administration during the initialcourse is such that there is more than 168 hours gap betweenadministration of two doses. In some cases, the initial course oftreatment comprises about 1 week and the frequency of administration isonce daily. In some cases, the initial course of treatment comprisesabout 1 week and the frequency of administration is such that there ismore than 24 hours gap between administration of two doses. In somecases, the initial course of treatment comprises about 1 week and thefrequency of administration is such that there is more than 48 hours gapbetween administration of two doses. In some cases, the initial courseof treatment comprises about 1 week and the frequency of administrationis such that there is more than 72 hours gap between administration oftwo doses. In some cases, the initial course of treatment comprisesabout 1 week and the frequency of administration is such that there ismore than 96 hours gap between administration of two doses. In somecases, the initial course of treatment comprises about 1 week and thefrequency of administration is such that there is more than 120 hoursgap between administration of two doses. In some cases, the initialcourse of treatment comprises about 1 week and the frequency ofadministration is such that there is no more than 168 hours gap betweenadministration of two doses. In some cases, the initial course oftreatment comprises about 1 week and the frequency of administration issuch that one dose is administered during the week. In some examples,the initial course of treatment comprises about 2 weeks and thefrequency of administration is such that one dose is administered duringthe 2 weeks. In some cases, the initial course of treatment comprisesabout 2 weeks and the frequency of administration is such that at leasttwo doses are administered during the 2 weeks. In some cases, theinitial course of treatment comprises about 3 weeks and the frequency ofadministration is such that only one dose is administered during thethree weeks.

In some embodiments, the composition is intravaginally administeredduring the subsequent course of treatment daily, every other day, once aweek, once every 2 weeks, once every 3 weeks, once every 4 weeks, onceevery 5 weeks, once every 6 weeks, once every 7 weeks, once every 8weeks, once every 9 weeks, once every 10 weeks, once every 11 weeks,once every 12 weeks, once every 13 weeks, once every 14 weeks, onceevery 15 weeks, once every 16 weeks, once every 17 weeks, once every 18weeks, or once every 19 weeks. In some embodiments, the frequency ofadministration during the subsequent course is every other day, once aweek, or twice a week. In some cases, the subsequent course of treatmentcomprises about 1 week and the frequency of administration is less thanonce daily. In some embodiments, the frequency of administration duringthe subsequent course is such that there is more than 24 hours gapbetween administration of two doses. In some embodiments, the frequencyof administration during the subsequent course is such that there ismore than 48 hours gap between administration of two doses. In someembodiments, the frequency of administration during the subsequentcourse is such that there is more than 72 hours gap betweenadministration of two doses. In some embodiments, the frequency ofadministration during the subsequent course is such that there is morethan 96 hours gap between administration of two doses. In someembodiments, the frequency of administration during the subsequentcourse is such that there is more than 120 hours gap betweenadministration of two doses. In some embodiments, the frequency ofadministration during the subsequent course is such that there is morethan 168 hours gap between administration of two doses. In some cases,the subsequent course of treatment comprises about 1 week and thefrequency of administration is less than once daily. In some cases, thesubsequent course of treatment comprises about 1 week and the frequencyof administration is such that there is more than 24 hours gap betweenadministration of two doses. In some cases, the subsequent course oftreatment comprises about 1 week and the frequency of administration issuch that there is more than 48 hours gap between administration of twodoses. In some cases, the subsequent course of treatment comprises about1 week and the frequency of administration is such that there is morethan 72 hours gap between administration of two doses. In some cases,the subsequent course of treatment comprises about 1 week and thefrequency of administration is such that there is more than 96 hours gapbetween administration of two doses. In some cases, the subsequentcourse of treatment comprises about 1 week and the frequency ofadministration is such that there is more than 120 hours gap betweenadministration of two doses. In some cases, the subsequent course oftreatment comprises about 1 week and the frequency of administration issuch that there is no more than 168 hours gap between administration oftwo doses. In some cases, the subsequent course of treatment comprisesabout 1 week and the frequency of administration is such that one doseis administered during the week. In some examples, the subsequent courseof treatment comprises about 2 weeks and the frequency of administrationis such that one dose is administered during the 2 weeks. In some cases,the subsequent course of treatment comprises about 2 weeks and thefrequency of administration is such that at least two doses areadministered during the 2 weeks. In some cases, the subsequent course oftreatment comprises about 3 weeks and the frequency of administration issuch that only one dose is administered during the three weeks.

In some embodiments, administration of the composition described hereinusing methods described herein results in lowering of vaginal pH. Insome embodiments, lowering of vaginal pH is observed after the firstadministration of the composition during a course of treatment. In someembodiments, lowering of vaginal pH is observed after at least twoadministrations of the composition during a course of treatment. In someembodiments, lowering of vaginal pH is observed after multipleadministrations of the composition during a course of treatment. In someembodiments, lowering of vaginal pH is observed after the initial courseof treatment. In some embodiments, lowering of vaginal pH is observedafter the subsequent course of treatment. In some instances,administration of the composition lowers the vaginal pH by at least 0.1compared to the baseline vaginal pH prior to the administration of thecomposition. In some instances, administration of the composition lowersthe vaginal pH by at least 0.2 compared to the baseline vaginal pH priorto the administration of the composition. In some instances,administration of the composition lowers the vaginal pH by at least 0.3compared to the baseline vaginal pH prior to the administration of thecomposition. In some instances, administration of the composition lowersthe vaginal pH by at least 0.4 compared to the baseline vaginal pH priorto the administration of the composition. In some instances,administration of the composition lowers the vaginal pH by at least 0.5compared to the baseline vaginal pH prior to the administration of thecomposition. In some instances, administration of the composition lowersthe vaginal pH by at least 0.6 compared to the baseline vaginal pH priorto the administration of the composition. In some instances,administration of the composition lowers the vaginal pH by at least 0.7compared to the baseline vaginal pH prior to the administration of thecomposition. In some instances, administration of the composition lowersthe vaginal pH by at least 0.8 compared to the baseline vaginal pH priorto the administration of the composition. In some instances,administration of the composition lowers the vaginal pH by at least 0.9compared to the baseline vaginal pH prior to the administration of thecomposition. In some instances, administration of the composition lowersthe vaginal pH by at least 1.0 compared to the baseline vaginal pH priorto the administration of the composition. In some instances,administration of the composition lowers the vaginal pH by about 0.1compared to the baseline vaginal pH prior to the administration of thecomposition. In some instances, administration of the composition lowersthe vaginal pH by about 0.2 compared to the baseline vaginal pH prior tothe administration of the composition. In some instances, administrationof the composition lowers the vaginal pH by about 0.3 compared to thebaseline vaginal pH prior to the administration of the composition. Insome instances, administration of the composition lowers the vaginal pHby about 0.4 compared to the baseline vaginal pH prior to theadministration of the composition. In some instances, administration ofthe composition lowers the vaginal pH by about 0.5 compared to thebaseline vaginal pH prior to the administration of the composition. Insome instances, administration of the composition lowers the vaginal pHby about 0.6 compared to the baseline vaginal pH prior to theadministration of the composition. In some instances, administration ofthe composition lowers the vaginal pH by about 0.7 compared to thebaseline vaginal pH prior to the administration of the composition. Insome instances, administration of the composition lowers the vaginal pHby about 0.8 compared to the baseline vaginal pH prior to theadministration of the composition. In some instances, administration ofthe composition lowers the vaginal pH by about 0.9 compared to thebaseline vaginal pH prior to the administration of the composition. Insome instances, administration of the composition lowers the vaginal pHby about 1.0 compared to the baseline vaginal pH prior to theadministration of the composition.

In some embodiments, administration of the composition described hereinusing methods described herein restores a healthy vaginal microbiome. Insome embodiments, restoration of a healthy vaginal microbiome ischaracterized by an increase in population of aerobic microbes and adecrease in population of anaerobic microbes, such as Gardnerellavaginalis, Prevotella, Peptostreptococcus, and Bacteroides spp. In someembodiments, a healthy vaginal microbiome is restored after the firstadministration of the composition during a course of treatment. In someembodiments, a healthy vaginal microbiome is restored after at least twoadministrations of the composition during a course of treatment. In someembodiments, a healthy vaginal microbiome is restored after the initialcourse of treatment. In some embodiments, a healthy vaginal microbiomeis restored after the subsequent course of treatment.

In some embodiments, the compositions of the present disclosure areadministered to the vagina of the mammal in a dosage comprising about0.5 g to about 10 g, about 2 g to about 9 g, about 3 g to about 8 g,about 4 g to about 7 g, about 5 g to about 6 g, about between 3 g toabout 4 g, or about 4 g to about 5 g of the composition. The dosage ofadministration, in some embodiments comprises about 0.5 g, about 0.6 g,about 0.7 g, about 0.8 g, about 0.9 g, about 1 g, about 1.1 g, about 1.2g, about 1.3 g, about 1.4 g, about 1.5 g, about 1.6 g, about 1.7 g,about 1.8 g, about 1.9 g, about 2 g, about 2.1 g, about 2.2 g, about 2.3g, about 2.4 g, about 2.5 g, about 2.6 g, about 2.7 g, about 2.8 g,about 2.9 g, about 3 g, about 3.1 g, about 3.2 g, about 3.3 g, about 3.4g, about 3.5, about 3.6 g, about 3.7 g, about 3.8, about 3.9, about 4 g,about 5 g, about 5.1 g, about 5.2 g, about 5.3 g, about 5.4 g, about 5.5g, about 5.6 g, about 5.7 g, about 5.8 g, about 5.9 g, about 6 g, about7 g, about 7.1 g, about 7.2 g, about 7.3 g, about 7.4 g, about 7.5 g,about 7.6 g, about 7.7 g, about 7.8, about 7.9 g, about 8 g, about 8.1g, about 8.2 g, about 8.3 g, about 8.4 g, about 8.5 g, about 8.6 g,about 8.7 g, about 8.8 g, about 8.9 g, about 9 g, about 9.1 g, about 9.2g, about 9.3, about 9.4 g, about 9.5 g, about 9.6 g, about 9.7, about9.8 g, about 9.9 g, about 10 g, about 10.1 g, about 10.2 g, about 10.3g, about 10.4 g, about 10.5 g, about 10.6 g, about 10.7 g, about 10.8 g,about 10.9 g, about 11 g, about 11.1 g, about 11.2 g, about 11.3 g,about 11.4 g, about 11.5 g, about 11.6 g, about 11.7 g, about 11.8 g,about 11.9 g, about 12 g, about 12.1 g, about 12.2 g, about 12.3 g,about 12.4 g, about 12.5 g, about 12.6 g, about 12.7 g, about 12.8 g,about 12.9 g, about 13 g, about 13.1 g, about 13.2 g, about 13.3 g,about 13.4 g, about 13.5 g, about 13.6 g, about 13.7 g, about 13.8 g,about 13.9 g, about 14 g, about 14.1 g, about 14.2 g, about 14.3 g,about 14.5 g, about 14.6 g, about 14.7 g, about 14.8 g, about 14.9 g,about 15 g, about 15.1 g, about 15.2 g, about 15.3 g, about 15.4 g,about 15.5 g, about 15.6 g, about 15.7 g, about 15.8 g, about 15.9 g,about 16 g, 16.1 g, about 16.2 g, about 16.3 g, about 16.4 g, about 16.5g, about 16.6 g, about 16.7 g, about 16.8 g, about 16.9 g, about 17 g,about 17.1 g, about 17.2 g, about 17.3 g, about 17.4 g, about 17.5 g,about 17.6 g, about 17.7 g, about 17.8 g, about 17.9 g, about 18 g,about 18.1 g, about 18.2 g, about 18.3 g, about 18.4 g, about 18.5 g,about 18.6 g, about 18.7 g, about 18.8 g, about 18.9 g, about 19 g,about 19.1 g, about 19.2 g, about 19.3 g, about 19.4 g, about 19.5 g,about 19.6 g, about 19.7 g, about 19.8 g, about 19.9 g, or about 20 g ofthe composition.

The present disclosure further provides a method for prognosis of riskof recurrence of BV, the method comprising assessment of BV in a subjectby testing vaginal pH, administration of the composition describedherein according to the methods described herein, followed byre-assessment of BV in the subject by re-testing vaginal pH, andprognosing the risk of recurrence of BV based of comparisons of vaginalpH before and after administration of the composition. In someembodiments, the vaginal pH is lowered after administration of thecomposition described herein according to methods described above. Insome embodiments, lowering of vaginal pH in a subject followingadministration of the composition described herein according to themethods described herein is associated with good prognosis for reducedrisk of recurrence of BV in the subject.

Another embodiment provides a method for prognosis of risk of recurrenceof BV, the method comprising assessment of BV in a subject by a firstvaginal microbiome analysis, administration of the composition describedherein according to the methods described herein, followed byre-assessment of BV in the subject by a subsequent vaginal microbiomeanalysis, and prognosing the risk of recurrence of BV based ofcomparisons of vaginal microbiome analysis before and afteradministration of the composition. In some embodiments, administrationof the composition described herein according to methods describedabove, restores a healthy vaginal microbiome in a subject. In someembodiments, restoration of a healthy vaginal microbiome ischaracterized by an increase in population of aerobic microbes, such asLactobacillus spp. and a decrease in population of anaerobic microbes,such as Gardnerella vaginalis, Prevotella, Peptostreptococcus, andBacteroides spp. In some embodiments, restoration of a healthy vaginalmicrobiome in a subject following administration of the compositiondescribed herein according to the methods described herein is associatedwith good prognosis for reduced risk of recurrence of BV in the subject.

A further embodiment provides a method of prognosis of risk ofrecurrence of BV, the method comprising assessment of BV in a subjectusing Amsel criteria, administration of the composition described hereinaccording to the methods described herein, followed by re-assessment ofBV in the subject using Amsel criteria, and prognosing the risk ofrecurrence of BV based of comparisons of Amsel criteria results beforeand after administration of the composition. In some embodiments,following intravaginal administration of the composition the subjectdoes not meet at least three of four Amsel criteria. The subject is thendefined as Amsel-negative. In some embodiments, the subject isAmsel-negative after a first intravaginal administration of thecomposition during a course of treatment. In some embodiments, thesubject is Amsel-negative after at least two intravaginaladministrations of the composition during a course of treatment. In someembodiments, the subject is Amsel-negative after the initial course oftreatment. In some embodiments, the subject is Amsel-negative after thesubsequent course of treatment. In some embodiments, an Amsel-negativesubject is considered to have a good prognosis for reduced risk ofrecurrence of BV in the subject. In some embodiments, an Amsel-positivesubject is considered to have a poor prognosis for reduced risk ofrecurrence of BV in the subject.

EXAMPLES

The following specific, non-limiting examples are to be construed asmerely illustrative, and do not limit the present disclosure of thescope of the disclosure. Without further elaboration, it is believedthat one skilled in the art can, based on the description herein,utilize the present disclosure to its fullest extent.

Example 1: A Randomized, Placebo Controlled Pilot Study to Determine theEffect and Duration of One Exemplary Gel Formulation (e.g., Acidform orAmphora® Gel) on Vaginal pH

A total of 100 female subjects, aged 18 to 45 years, will be enrolledfor the study. The study will be carried out in two sites with 50subjects for each site. It is anticipated there will be approximately 26weeks, including a three month period to complete enrollment, betweenstudy activation and the final monitoring close-out visit. The enrolledsubjects will participate in the study according to the followingschedule: Screening, Day 0; Admission/Dosing/Monitoring, Day 1;Discharge, Day 2; Self-obtained vaginal pH readings, Days 2-6;Outpatient clinic visit, Day 7.

An intravaginally (IVAG) applied exemplary gel formulation (e.g.,Acidform or Amphora® Gel) will be used as the investigational product(IP) in the study. The exemplary gel formulation (e.g., Acidform orAmphora® Gel) is an acidity-maintaining gel (pH 3.5) containing threeacidic compounds (citric acid, potassium bitartrate, and lactic acid), apreservative (benzoic acid), two natural polymer thickeners (alginicacid and xanthan gum), a humectant (glycerin), and water. The exemplarygel formulation (e.g., Acidform or Amphora® Gel) will be administered ata dosage of 3, 4, or 5 g, IVAG.

Universal Placebo Gel (UPG), an isotonic non-buffering gel, pH adjustedto 4.5, containing 2.7% hydroxyethylcellulose, sorbic acid, sodiumhydroxide, sodium chloride and purified water, will be used as acontrol. UPG will be administered at a dosage of 4 g, IVAG.

Primary Objective of the study is to determine effect and duration ofone IVAG dose (3, 4 or 5 g) of the exemplary gel formulation (e.g.,Acidform or Amphora® Gel) or placebo gel (4 g UPG) or no treatment, onvaginal pH in women.

Exploratory objectives of the study are: (1) To determine the effect ofone IVAG dose of the exemplary gel formulation (e.g., Acidform orAmphora® Gel) at a dose of 3, 4 or 5 g, or placebo gel (4 g UPG) or notreatment, on asymptomatic bacterial vaginosis (BV) based on Amselcriteria, (2) To determine the effect of a single application of theexemplary gel formulation (e.g., Acidform or Amphora® Gel) at a dose of3, 4 or 5 g on the vaginal microbiome.

Primary Outcome measures of the study is to assess change in vaginal pHand duration of change from baseline pH following a single dose (3, 4 or5 g) of WAG the exemplary gel formulation (e.g., Acidform or Amphora®Gel), placebo gel (4 g UPG), or no treatment. Exploratory outcomemeasures: (1) To determine the effect of a single dose (3, 4 or 5 g) ofIVAG the exemplary gel formulation (e.g., Acidform or Amphora® Gel),placebo gel (4 g UPG) or no treatment on asymptomatic BV based onAmsel's criteria, (2) To determine the effect of a single application ofthe exemplary gel formulation (e.g., Acidform or Amphora® Gel) at a doseof 3, 4 or 5 g on the vaginal microbiome.

Study Participants: One hundred volunteer women, 20 per each treatmentarm, will be treated with either IVAG the exemplary gel formulation(e.g., Acidform or Amphora® Gel), 5 g dose (GROUP A); 4 g dose (GROUPB); 3 g dose (GROUP C); or Universal Placebo Gel (UPG), 4 g (GROUP D);or no treatment (GROUP E). For GROUPS A, B, C and D, treatment isdefined as speculum exam plus instillation of Investigational Product(IP) or placebo gel. For GROUP E (control), treatment is defined asspeculum exam and no gel instillation. At least 13 women in each groupwill be of either African American or Hispanic ethnicity.

Study Protocol Routine screening tests will be performed on admission;and subjects will be assessed for asymptomatic BV via vaginal swabsobtained for grading by Amsel criteria. A direct vaginal pH reading willbe research staff-obtained before the speculum exam, as well as onehour, and six hours post-treatment (Day 0). At the one hour and sixhours post treatment time points, direct vaginal pH readings will betaken on specimens collected from two different positions in the vagina,in case of incomplete distribution of the IP immediately followinginstillation. Both readings will be included as data points. At the sixhours time point, subjects will be trained on self-collecting vaginalswabs and performing the vaginal pH test. At 12 hours post-treatment,subjects will perform the vaginal pH test themselves using self-obtainedswabs, and record their results for clinician review. Clinicians willcollect AE assessment data will at each post treatment vaginal pHtesting time point while subjects are in the domiciliary unit (1, 6, 12,and 24 hours post-treatment).

The vaginal microbiome collection will be done on the women who consentfor this. The same swab used for vaginal pH at the Baseline/Screeningwill be used for the microbiome analysis post 24 hours treatment and ondays 2 through 7. The research staff will obtain the Baseline/Screening(a repeat pH/microbiome swab will be taken if screening and admission tothe domiciliary unit does not take place on the same day), post 24 hourtreatment and day 7 readings. The subjects will obtain the microbiomereadings on days 2-6 at home, concurrently with the pH readings.

Subjects will stay overnight in the domiciliary unit of the clinicaltrial center, and vaginal pH, microbiome collection (consented subjects)and Amsel criteria will be measured again by research staff at 24 (+/−2)hours post-treatment before discharge on Day 1. The subjects will bedischarged with the appropriate pH testing supplies and diary. All womenmust agree to abstain from sexual intercourse, douching and use of anyintravaginally applied products or devices until after their final studyvisit on Day 7. Subjects will measure their vaginal pH and do themicrobiome collection (if consented) at 24 (+/−4) hour intervals for 5days (Days 2-6) as outpatients and record the pH test results, anychange in vaginal comfort, and confirmation of microbiome swab saved(for consented subjects) in a provided diary. Subjects will also recordany activities engaged in from the abstinence criteria in the studyexclusion list, if applicable, each day. On Day 7 (+/−24 hours),subjects will return to the clinic with their diaries, swab transportcontainers, have their vaginal pH and Amsel criteria determined by theclinic staff, and queried as to any vaginal discomfort (vaginal comfortassessment) over the course of the study, as self-recorded in theirdiaries.

Screening and Eligibility: Screening, eligibility assessment andenrollment may all occur on the same day. Admission to the domiciliaryunit for dosing and 24 hour observation may also occur on the same dayas the Screening visit.

However, enrolled subjects who are unable to be admitted for dayactivities on the same day as screening may be admitted up to 5 dayspost screening, provided the following: (1) Subjects who are notadmitted on the same day as screening, but within 48 hours of screening,have a urine pregnancy (UHCG) and vaginal pH test, medical/surgicalhistory, concomitant medications, and vital signs repeated uponadmission. If subject has consented to microbiome collection, the swabwill again be collected, (2) Subjects admitted more than 48 hours but upto 5 days post Screening have a urine pregnancy (UHCG), vaginal pH test,speculum exam, vaginal smear for Amsel criteria, medical/surgicalhistory, concomitant medications, and vital signs repeated uponadmittance. If subject consents to microbiome collection, the swab willagain be collected.

Intervention and Follow-up Period: The Intervention and Follow-up Periodis Day 0-Day 7 in the Schedule of Events (Appendix A). The protocoldefined period of observation is 8 days.

Safety Follow-up After Protocol Defined Period of Observation: Thisperiod includes safety follow-up as required by Good Clinical Practiceand standard clinical trial practice. Adverse events continuing afterDay 7 (Final Study Visit) will be followed until resolution or stablestatus as determined by the principal investigator at the clinical trialsite.

Study Outcome Measures:

Primary Outcome Measures will include assessment of change in vaginal pHfrom baseline, post-instillation of one dose of IVAG the exemplary gelformulation (e.g., Acidform or Amphora® Gel), placebo gel, or notreatment; as well as duration of this change in vaginal pH.

Exploratory Outcome Measures will include assessment of effect of asingle dose of IVAG the exemplary gel formulation (e.g., Acidform orAmphora® Gel) gel, placebo gel or no treatment on asymptomatic BV, basedon Amsel criteria.

Study Population: Approximately 150 women will be screened to identify100 female subjects who are eligible to participate. Up to fifteenadditional subjects will be selected to account for subjects who arelost to follow up or who are withdrawn.

Schedule for individual subjects: Individual subjects will participatein the study for approximately eight days. The study schedule to befollowed is shown in FIG. 1 and also described below in details.

Study enrollment and retention: No study procedures will be performeduntil informed consent is obtained as part of the enrollment process.One hundred female subjects, ages 18-45, will be enrolled. Subjects willbe recruited through IRB-approved advertisements, database queries, andword of mouth. Children, pregnant women, prisoners, and other vulnerablepopulations will not be enrolled. Because vaginal pH values have beenfound to vary by ethnicity, recruitment efforts will focus on enrollmentof at least 65 women of African American or Hispanic descent (ideally,nearly equal numbers of each race at each site).

Screening will begin with the principal investigator at the clinicaltriat site or a designee providing an overview of the study to thepotential subject. Subjects will be excluded if they do not understandthe protocol and participation requirements and/or if they are unlikelyor unwilling to remain in the domiciliary unit overnight, conductvaginal pH readings on self-obtained vaginal swabs at home, or attendthe Day 7 follow-up visit. Potential subjects will be invited to thestudy-designated clinic area for additional screening procedures andinformation about the study. Potential subject will be assigned ascreening number. Study retention strategies will include clearexplanations during enrollment of the study schedule and procedures andreimbursement to support travel and time spent in clinic. Subjects willbe reminded of visits beforehand, and study staff will contact those whomiss appointments.

Eligibility Criteria: All eligibility criteria must be met for inclusioninto the study; no waivers will be granted.

Inclusion Criteria:

-   -   1. Female subjects between 18 and 45 years, inclusive    -   2. Ability to understand the consent process and procedures    -   3. Subjects agree to be available for all study visits    -   4. Written informed consent in accordance with institutional        guidelines    -   5. Negative pregnancy test    -   6. Able and willing to comply with all study procedures    -   7. Have not engaged in sexual intercourse, douching or used of        any form of vaginal suppository or intravaginal device for 24        hours prior to enrollment.    -   8. Agree to abstain from sexual intercourse, douching or any        form of vaginal suppository or intravaginal device use during        course of study    -   9. Report menstrual cycle regularity (25- to 35- day menstrual        cycles)    -   10. Subjects who test negative for BV or are positive for BV,        but asymptomatic. (BV positive subjects will be referred for        treatment at or following the Day 7 follow-up visit)

Exclusion Criteria:

-   -   1. Participation in any study with an investigational compound        or device within 30 days prior to signing informed consent    -   2. Active drug or alcohol use or dependence that, in the opinion        of the investigator, would interfere with adherence to study        protocol    -   3. Any other medical condition(s) that, in the judgment of the        investigator, might interfere with the study or require        treatment that might interfere with the study    -   4. Family member of the investigation study staff    -   5. Pregnant or breast-feeding    -   6. Inability to provide informed consent    -   7. A subject with a history or expectation of noncompliance with        medications or treatment protocol    -   8. Women with symptoms of UTI, symptomatic BV, yeast infection        or STI reported or observed during examination or based on        laboratory testing performed. (Should the study clinician make a        presumptive diagnosis of such infections, these subjects will be        withdrawn, referred for standard of care treatment, but will not        be followed as part of this study. They may be re-screened for        potential enrollment 14 days after treatment per Section 6.3,        Prohibited Medications, if all other screening criteria are        met.)    -   9. Women who regularly use douches, vaginal medications or        suppositories, feminine sprays, genital wipes or contraceptive        spermicides, or report abnormal vaginal discharge in the past 48        hours prior to screening    -   10. Women who are menstruating or who would expect to menstruate        during the study    -   11. Women who are currently using contraceptives that are        directly delivered to the vaginal mucosa, such as NuvaRing    -   12. Any specific condition that, in the judgment of the        Investigator, precludes participation because it could affect        subject safety

Prohibited Medications: Subjects must not be currently taking orapplying, or have taken or applied, for a period of 14 days prior toscreening, any antibiotic, antimycotic, or probiotic compounds, oral orintravaginal.

Randomization: This is a Phase I randomized controlled trial, 20subjects per treatment arm; for a total of 100 subjects. Up to 15alternates may be enrolled to account for study subjects that drop outor are lost to follow-up. The 100 subjects will be randomized in a1:1:1:1:1 fashion across the five study groups. Simple randomizationwill be performed and all documentation of this procedure and outputwill be saved with the study biostatistician's files until the end ofthe study. The randomization list will be generated by the studybiostatistician and transferred to study pharmacist prior to start ofthe study. Randomization will occur following enrollment and prior toDay 0 procedures. Alternates will be screened and enrolled if randomizedsubjects do not meet inclusion criteria on Day 0.

Blinding: Study subjects in Groups A, B, C and D will remain blinded totheir treatment assignment (with the exemplary gel formulation (e.g.,Acidform or Amphora® Gel) at 5 g, 4 g, 3 g, or placebo gel), during theentire duration of the study. The Sponsor, Evofem, Inc., will prepare IPas well as placebo, and supply these products in ready to dispensestatus.

Withdrawal and Reasons for Withdrawal: A study subject will bediscontinued from participation in the study for:

-   -   Development of any Exclusion criteria;    -   Pregnancy or breastfeeding;    -   Request by subject to terminate participation;    -   Requirement for prohibited treatment (see Exclusion criteria)        before Day 7    -   Treatment-related toxicity;    -   Failure to adhere to requirements of the protocol including        treatment and safety monitoring;    -   Lost to follow-up;    -   Request of primary care provider;    -   At the request of the IRB/Ethics committee, FDA, or the Sponsor;    -   Incarceration;    -   The subject's well-being, based on the opinion of the clinical        trial site principal investigator.

Handling of Withdrawal: If a subject is withdrawn from participation,the reason(s) for discontinuation must be documented in the sourcedocuments and eCRFs. Subjects who withdraw or are withdrawn from thestudy who received any amount of the study product will be encouraged toreturn for a follow-up (with subjects' consent) visit for safety. Thesubject will be asked for consent to complete an end-of-study evaluation(all procedures from Final Study Visit, Day 7). If an AE or SAE hasoccurred, every effort will be made to undertake protocol-specifiedsafety follow-up procedures, and the subject will be encouraged toreceive appropriate care under medical supervision until the symptoms ofany AE resolve or the subject's condition becomes stable.

Handling Lost to Follow-up: The study design has incorporated primary,secondary, and tertiary strategies for reducing lost to follow-up.Primary strategies include having sites: fully inform subjects of studyvisit details prior to randomization, addressing subjects' expectationson study participation, motivating subjects to adhere to follow-upvisits and research protocols, tracking subjects with overdue visits andhaving an alternate subject contact on file. Secondary strategiesinclude: scheduling the Final Study Visit (Day 7) around subjectsavailability which are within the visit window, providing a visitreminder ahead of time, and reviewing data for missed and overduefollow-up visits. Tertiary strategies include: discussing subjects'difficulty in adhering to study protocol and answering any question theymight have, continuing attempts to reach lost to follow-up subject andrecording all contact attempts. Documentation of lost to follow-upstatus will be maintained in the subject's CRF and in source documents.The reason(s) for the lost to follow-up, if known, and the appropriatetimelines for subject-study site correspondence will be captured alongwith the study site's attempts to reach the subject. Lastly, all properprocedures of sending certified letter of discontinuation from studybased on lost to follow-up status and a recommendation to come in for adiscontinuation visit to determine safety of the subject will befollowed by the study site.

Alternate Subject Enrollment to Account for Lost to Follow-ups andWithdrawals: If an enrolled subject is lost to follow-up or withdrawalas described in Section 6.6.3 at any point prior to and including Day 7(Final Study Visit) the following procedure will apply. An alternatesubject who is enrolled in the study to account for a subject whowithdrew from the study, or was Lost to Follow-up, will be assigned fromthe alternates list, and randomized per the study's randomizationprocedures.

Termination of the Study: The study may be terminated at the discretionof the Sponsor for concerns of subject safety or any other reason. Thestudy may also be closed by the clinical trial sites with the agreementof the sponsor, the FDA, or as the result of a natural disaster.

Investigational product (IP): The IP, which is one exemplary gelformulation (e.g., Acidform or Amphora® Gel) is an acidity-maintaininggel (pH 3.5) containing three acidic compounds (citric acid, potassiumbitartrate, and lactic acid), a preservative (benzoic acid), two naturalpolymer thickeners (alginic acid and xanthan gum), a humectant(glycerin), and water. Sodium hydroxide is used to adjust the pH duringmanufacture. The placebo used in this study is Universal Placebo Gel(UPG). UPG is an isotonic non-buffering gel, pH adjusted to 4.5,containing 2.7% hydroxyethylcellulose, sorbic acid, sodium hydroxide,sodium chloride and purified water. In clinical trials, UPG has beenproven safe and acceptable when used up to twice daily for 14 days.Table 1 lists the ingredients of the exemplary formulation, IP, that wasused in the clinical trial, for which results are provided in Example 2.

TABLE 1 Formulation Chart for IP Ingredient % w/w Benzoic Acid 0.20 10NNaOH 2.20 Purified Water USP 74.00 Potassium Bitartrate 0.40 Citric Acid1.00 Lactic Acid 90% Solution 2.00 Alginic Acid (Protacid F120) 4.25Glycerin 8.00 Xanthan Gum (Vanzan NF-C) 3.00 10N NaOH adjust to pH 3.5As needed Purified Water USP Qs to 100

Formulation, and Packaging of the IP: For this study, single useapplicators will be used. The IP and placebo gels will be filled intounit-dose applicators consisting of an injection molded opaque whitepolyethylene barrel with a rounded distal end for easy insertion. Athermoplastic rubber piston will be inserted at the opposite end of thebarrel. This component will seal the end of the barrel to keep thecontents stable over time and also functions as the applicator pistonwhen in use. A polyethylene plunger rod will be packaged alongside ofthe barrel within a plastic film sanitary overwrap. The research studykit will contain a group of overwrapped applicators placed into astandard white cardboard box along with instructions for use.

Product Supply: The IP, which is one exemplary gel formulation (e.g.,Acidform or Amphora® Gel) and the UPG placebo will be obtained from theSponsor.

Product Storage, Stability, and Expiration: The filled applicators willbe stored at room temperature (15° C.-30° C.). Applicators will bemaintained in the research pharmacy and dispensed by the site ResearchPharmacist.

Preparation: The study product is supplied ready for use. No preparationwill be necessary.

Administration: The study product will be administered (instilled) bythe study clinician on Day 0, following subject admission to thedomiciliary unit. Gel instillation occurs after the gynecologic exam; nospeculum is used during administration. In use, the plunger rod will beinserted into the piston end of the barrel engaging the piston. The capwill be removed from the distal end of the barrel; the barrel will thenbe inserted into the subject's vagina (distal end first). The plungerrod will be pushed into the barrel, subsequently moving the piston andgel toward the open end of the barrel. Only the gel will leave thebarrel. These are the instructions supplied to the research staff withthe study product.

TABLE 2 Investigational Product Dosing Treatment Group InvestigationalProduct* Placebo** Group A 20 subjects, 5 g dose N/A Group B 20subjects, 4 g dose N/A Group C 20 subjects, 3 g dose N/A Group D N/A 20subjects, UPG, 4 g dose Group E*** N/A N/A

The dosage of IP is 5 g, 4 g or 3 g (as described for each treatmentgroup) in a pre-filled applicator. Administered by clinician on Day 0only

The dosage of placebo (UPG) is 4 g in a pre-filled applicator.Administered by clinician on Day 0 only. Control group will receive notreatment.

Accountability/Final Disposition for the Investigational Product(s): Theclinical trial site principal investigator will keep a record of thedates and amounts of IP and placebo received, including packing slips,the amount dispensed to study subjects, and the amount unused. Theserecords should include the dates, quantities, batch/serial numbers andexpiration dates (if applicable).

The Site PI will record drug accountability on a Drug Disposition Log.All used applicators can be discarded per clinic policy (Appendix D).Upon completion of the study and the final drug accountabilitymonitoring visit, all unused study medication will be returned to thesponsor or, if instructed by the sponsor, unused study product may bedestroyed onsite according to the site Research Pharmacy procedures.

Detailed study procedures/evaluations: Subjects will undergo thefollowing study procedures as indicated in this section. All equipmentused will be calibrated and validated as appropriate.

Written Informed Consent: A written Informed Consent will be obtainedfrom all study participants before start of any study activities. TheInformed Consent has been developed in accordance with the InternationalConference on Harmonization Good Clinical Practices (ICH-GCP)guidelines. The Informed Consent specifies, in lay and culturallyappropriate language, all expectations from the participant includingduration of study, number of doses and visits, procedures at each visit,safety documentation including subject diary, explanation of abstinenceexpectations, restrictions on intravaginal products or device use,sampling plan, potential risks, stipend for participation in study, andrelevant research scope. All study-related questions by the subjectshould be responded to before completion of the Informed Consent. Thesubject will be provided ample time, privacy and any questions about theICF will be addressed. The completed Informed Consent form will beverified for its proper completion including that all required initialsand signatures are in place. A signed copy of the Informed Consent willbe provided to the subject and the original will be kept on-file in thesubject's study-file (documentation).

Inclusion/Exclusion Criteria: Participants will meet all the statedinclusion criteria and not possess any of the exclusion criteria to beeligible to participate in this study. Only subjects that are eligiblebased on the trial's inclusion and exclusion criteria will be enrolledin the trial.

Evaluation of Eligibility Criteria: The clinical trial site principalinvestigator will evaluate whether a subject meets the study'sinclusion/exclusion criteria as mentioned in the Inclusion/ExclusionCriteria listed above, at screening and before administering any studyproduct. The clinical trial site principal investigator will beresponsible for ensuring that the evaluation of eligibility criteria isaddressed for all subjects enrolled in the study.

Demographics: Demographics data will be collected from all studyparticipants. Subject's date of birth, race, and ethnicity will becollected and recorded as part of demographics data.

Clinical Evaluations: Summarized procedural steps for these evaluationsare provided for information only; all procedures will be performedaccording to site/clinic standard of care SOPs (standard operatingprocedures).

Medical History: A detailed medical/surgical/gynecologic/sexual healthhistory will be obtained by a trained study staff. Changes in medicalhistory, if applicable, will be reviewed at follow-up visits. A reviewof body systems will be performed by the principal investigator at theclinical trial site (Site PI) to screen for exclusion criteria.

Vital Signs: Vital signs, including blood pressure, heart rate, and bodytemperature will be measured and recorded. Vital sign measurements willbe obtained after the subject has been sitting quietly for at least 5minutes. When taking an oral temperature, assure the subject has not hadany recent hot or cold beverages or smoking. The Site PI should useclinical judgment when characterizing bradycardia among health subjects,for example conditioned athletes.

Gynecologic Examinations: A full gynecologic examination (speculum) willbe performed by a licensed practitioner, after swabs for vaginal pH andmicrobiome collection (in consented subjects) are collected. Clinicianswill follow clinic procedures for these examinations, which in generalwill be performed as follows:

-   -   1. Each step of the exam will be explained to the subject    -   2. The subject is asked to assume the lithotomy position    -   3. External genitalia are assessed for abnormalities; i.e, signs        of infection or inflammation    -   4. The closed speculum is introduced at a downward 45° angle.        Should lubrication be necessary, warm water (only) may be used        to facilitate insertion of the speculum; use of water to        lubricate is noted in the source documents. After the speculum        has entered the vagina, the blades of the speculum are rotated        into a horizontal position. After full insertion, the speculum        blades are opened and maneuvered gently so that the cervix comes        into full view. The cervical os and vaginal walls are visually        inspected for presence of discharge, inflammation, or irritation

A targeted physical and/or gynecologic examination (speculum), will beperformed at any subsequent or unscheduled visit if necessary due to anadverse event (AE).

Concomitant Medications: Concomitant medications will be assessed andincludes any medication that would be taken during the study includingall over the counter medications, vitamins and nutritional supplements.The information collected for each concomitant medication included at aminimum: start date, stop date or continuing and indication.

Height and Weight: Body weight without coat and shoes and height withoutshoes will be recorded.

Laboratory Evaluations

Pregnancy Testing: Women of reproductive potential including those withhistory of tubal ligation participating in the study will be required tocollect a urine specimen to be tested for urine (3-human chorionicgonadotropin (UHCG). Negative results must be obtained prior to dosing.Pregnancy testing will not be required for women who have undergone atotal hysterectomy with clinical proof of the procedure performed(should be >2 months before start of study). Women who test positive forpregnancy will be excluded from participating in this study and bereferred for follow-up as per clinic protocol, not as part of thisstudy.

HIV Testing: All subjects will have blood drawn for a serum HIV test.Specimen collection and testing shall be performed in accordance withsite laboratory SOPs. HIV testing is performed for post-hoc analysis;HIV status is not an inclusion/exclusion criteria, therefore enrollmentof subjects will not be based on test results. Those subjects who testHIV positive will be referred for follow-up as per clinic protocol, notas part of this study. Subjects who have positive HIV laboratory resultsavailable in their medical record, will not need to be re-tested (thoseresults need to be placed in the research source record).

Vaginal pH Testing (Procedural steps for performing the vaginal pH testwill be supplied to research staff and subjects as a one page, step bystep pictorial reference sheet): Swab for vaginal pH reading will becollected prior to introduction of speculum for gynecologic exam.

Following instructions will be given to the research stuff for specimencollection:

-   -   1. Remove one strip of pH paper from the container. Place on a        sheet of plain white paper, in natural light.    -   2. Open swab (applicator) package, remove one swab, taking care        not to touch tip of swab to environmental or body surfaces.    -   3. Collect vaginal material on a swab:        -   a Insert one swab into the vagina approximately 2-3 inches,            and then rub the vaginal wall at this position in a circular            motion for approximately 15 seconds.        -   b. For sample collection while the subject is in the            domiciliary unit, each sampling time point will have up to 3            swabs collected (baseline, post treatment 1, 6, 12 and 24            hours). This sample is obtained as follows: Insert one swab            into the vagina and collect vaginal material from the apex            by rubbing the vaginal wall at this position in a circular            motion for approximately 15 seconds.    -   4. Microbiome collection in subjects who have consented:        -   a. Replace one of the pH collection swabs with the Copan            ESwab. Collect pH as above; following the placement of            vaginal secretion onto the pH paper, place swab into            transport medium container and store in freezer.

Following instructions will be given to the study participant forspecimen collection:

-   -   1. Remove one strip of pH paper; Place on a sheet of plain white        paper, in natural light.    -   2. Open swab (applicator) package, remove one swab, taking care        not to touch tip of swab to environmental or body surfaces.    -   3. Collect vaginal material on a swab by inserting one swab        approximately 2-3 inches into the vagina, and then rubbing the        vaginal wall in a circular motion for approximately 15 seconds.    -   4. For subjects that consented to the microbiome collection:        Place swab in transport medium container and store in freezer        until return to clinic on Day 7.

Following instructions will be given to the research stuff and studyparticipant for vaginal testing:

-   -   1. Remove swab from vagina and immediately, using a circular        motion, gently apply the vaginal material on the swab over the        surface of one strip of pH paper.    -   2 Immediately (within 30 seconds) observe the color reaction on        the paper and compare the color to the manufacturer supplied        color comparison chart.    -   3. Record the pH value corresponding to the color comparison        example closest in color on the comparison chart.

Following instructions will be given for Wet Prep Testing as a step bystep pictorial reference sheet:

-   -   1. Introduce speculum as described above in Gynecologic        Examinations section.    -   2. Open cotton tipped applicator package, remove one swab,        taking care not to touch tip of swab to environmental or body        surfaces.    -   3. Collect vaginal material on a swab by inserting one swab        approximately 2-3 inches into the vagina, and then rubbing the        vaginal wall in a circular motion for approximately 15 seconds,        or by collecting material from the posterior fornices. Note: If        sufficient material is collected, both the wet prep and the        whiff test can be performed on one swab.    -   4. Remove swab and place the collected material in a small test        tube containing approximately 0.5 mL of 0.85% nonbacteriostatic        normal saline (this sample can be used for both the whiff test        and the wet mount examination if necessary)    -   5. Label the tube with subject identifier    -   6. Maintain the specimen at room temperature (do not        refrigerate)    -   7. Apply one drop of collected material to a clean glass slide    -   8. Place a coverslip over the sample on the slide    -   9. Examine the slide at 10× and 40× objective for epithelial        cells, clue cells, budding yeast or pseudohyphae, and        trichomonads    -   10. Apply a coverslip over the slide with sample collected for        whiff test with KOH added. Make sure the KOH has had at least        30-60 seconds to react with the sample before observing    -   11. Examine the slide at 10× and 40× objective for budding        yeast, pseudohyphae and blastospores.

Wet prep is going to be considered positive for bacterial vaginosis ifclue cells are present; positive for vaginal candidiasis if buddingyeast or pseudohyphae are present; positive for trichomonas iftrichomonas parasites are present. (Should the study clinician make apresumptive diagnosis of such infections, these subjects will bewithdrawn and referred for standard of care treatment. They mayre-enroll and begin study activities 14 days after treatment per Sectiondescribing the Prohibited Medications.

Following instructions will be given for Whiff Amine Testing (KOH Test)(Summarized procedural steps for these tests are provided forinformation only; specimen collection and testing shall be performedaccording to site laboratory SOPs):

-   -   1. Apply a portion of the undiluted vaginal material from swab        collected in same manner as vaginal pH swab, or one drop of the        saline suspension (see Wet Prep Testing), to the surface of a        clean glass slide.    -   2. Add one drop of 10% potassium hydroxide (KOH) directly to the        vaginal specimen    -   3. Holding the slide gently fan the vapor layer (whiff) above        the surface of the slide and assess for the presence of volatile        amines which have a fishy odor.

Positive: The presence of a fishy odor following addition of KOH to thevaginal specimen. Negative: The absence of a fishy odor followingaddition of KOH to the vaginal specimen.

Amsel Criteria: Amsel criteria corresponding to the diagnosis of BV arelisted below and subject must be positive in three of these four areasto be diagnosed with bacterial vaginosis.

-   -   1. Vaginal pH>4.5    -   2. Presence of thin, grayish white vaginal discharge    -   3. Positive Whiff amine test    -   4. Positive Wet prep

Assessment of Vaginal Irritation (Vaginal Comfort): Prior to collectionof swabs for vaginal pH readings, all subjects will be asked to assesstheir current level of vaginal comfort. They will be asked to rate thisassessment using the following criteria and scale, listed in Table 3.

TABLE 3 Adverse events evaluation scale. Vaginal Comfort AssessmentCriteria Rating Vaginal pain None Mild Moderate Severe Vaginal burningNone Mild Moderate Severe Vaginal Itching None Mild Moderate SevereVaginal discharge None Mild Moderate Severe

While inpatients on Day 0 until discharge on Day 1, research staff willrecord vaginal comfort assessments in the source documents; for theself-performed 12 hour vaginal pH test and each day following discharge,subjects will record vaginal comfort assessments in their diary (i.e.,the Subject Diary), prior to collecting the swab to perform vaginal pHtesting. Subjects who rate any vaginal comfort criteria as “Moderate” orhigher will be assessed, (following discharge, subjects are instructedto report this information to the Site PI immediately) and ifapplicable, examined. All criteria assessed as “Mild” or higher will berecorded as an Adverse Event.

Subject Diary: Each subject will be provided with a subject diary totrack adverse events between study visits. Study Coordinator willexplain to each subject the importance of the diary. The instruction onmaintaining the diary should include what fields have to be completedand how the subject should record data within it. Study coordinator willreview the diary prior to discharge and at the final study visit. Thesubject will record vaginal pH readings (including date and time),vaginal comfort level, any adverse events, as well as noncompliance toactivities or medications noted in the subject exclusion list orconcomitant medications list at screening (abstaining from sexualintercourse, douching or any form of vaginal suppository or intravaginaldevice; any antibiotic, antimycotic, or probiotic compounds, oral orintravaginal, used during course of study). For subjects that haveconsented to the microbiome collection, they will record placement ofthe swab and its transport container into the freezer. Any deficienciesand attempts to correct these deficiencies should be noted in progressnotes (source documents). Study coordinator must ensure that the diariesare returned at the time designated in the study protocol. If a subjectdiary is not returned, the site should make several attempts to retrieveit. These attempts should be documented in the subject's sourcedocuments. The Subject diary will be reviewed by the research staffprior to each vaginal pH sample collection time point while subjects arein the domiciliary unit, and before start of visit-specific proceduresat the Day 7 Final Study visit. The diary will be issued to the subjecton Day 0, during training session to perform vaginal pH testing, andwill be kept by the subjects through Day 7 (Final Study Visit). Diarieswill be re-issued to the subject after making a copy for subjects' studyrecord. All returned diaries will become part of the subjects' studyrecord.

The following section describes the procedures to be carried out on eachvisit:

Day 0 (prior to admittance to domiciliary unit): Screening Enrollmentand admittance to domiciliary unit for dosing (Day 0) may all occur onthe same day. Once screened, eligible subjects must be enrolled andadmitted to the domiciliary unit for dosing (Day 0) within five (5) dayspost screening visit. Enrolled subjects who are not admitted within 48hours of Screening will have some tests/procedures repeated uponadmission. It is anticipated that about three subjects will be screenedfor every two enrolled. Subjects who wish to participate in the trialwill be asked to sign and date the Informed Consent and HIPAA Formsprior to any trial specific procedures. All subjects will be providedwith a copy of their own signed and dated Informed Consent Form. Theprincipal investigator at the clinical trial site must keep a subjectscreening log, subject enrollment log and subject identification log foridentifying all subjects having signed Informed Consent and HIPAA Forms.

The following will be performed by research staff and/or recorded in thesource documents and case report form (CRF):

-   -   1. Informed consent and HIPAA completed    -   2. Additional consent required for participating in the        Microbiome collection    -   3. Evaluation of inclusion and exclusion criteria    -   4. Evaluate eligibility criteria    -   5. Demographic data    -   6. Medical/surgical/gynecologic history    -   7. Concomitant medication: All medications taken will be        recorded in standard data collection forms with attention to        drug route, daily dose, duration, start date, stop date and        indications.    -   8. Body height and weight    -   9. Vital signs measurement (BP, HR, temperature)    -   10. Pregnancy test (UHCG)    -   11. Laboratory Testing (Vaginal pH, Microbiome collection, Wet        Mount Smear, Whiff test, all performed from vaginal swab—Amsel        criteria evaluation from results of these three tests)    -   12. Gynecologic examination (speculum, performed by clinician)    -   13. HIV serology (blood draw). If subject has a laboratory        result of HIV positive result, this blood draw does not need to        be done

Day 0 (at the clinical trial site domiciliary unit): Screening,Enrollment and admittance to domiciliary unit for dosing (Day 0) may alloccur on the same day. These procedures reflect a cross over from whatis completed for screening. The following will be performed and/orrecorded in the source documents and CRF:

-   -   1. Record any changes in the following since screening visit:        -   a. medical/surgical/gynecologic history        -   b. adverse events        -   c. concomitant medications    -   2. Vaginal comfort assessment (baseline)    -   3. Pregnancy test (urine)    -   4. Vaginal pH (performed by research staff)    -   5. Vaginal microbiome collection for subjects who consented    -   6. Gynecologic examination (speculum, performed by clinician)

If the subject withdraws or is terminated (for not meeting all theinclusion/exclusion criteria outlined above) from the study prior toadministration of the study product, the procedure described inAlternate Subject Enrollment to Account for Lost to Follow-ups andWithdrawals is followed.

Administration of IP or no treatment based on Group assignment.

-   -   1. GROUP A subjects: the exemplary gel formulation (e.g.,        Acidform or Amphora® Gel) at 5 g dose instilled IVAG    -   2. GROUP B subjects: the exemplary gel formulation (e.g.,        Acidform or Amphora® Gel) at 4 g dose instilled IVAG    -   3. GROUP C subjects: the exemplary gel formulation (e.g.,        Acidform or Amphora® Gel) at3 g dose instilled IVAG    -   4. GROUP D subjects: UPG, 4 g dose instilled IVAG    -   5. GROUP E subjects: No treatment (For GROUP E (control),        treatment is defined as no gel instillation. Time points for        subsequent procedures from Day 0 through Day 7 noted as “X hours        post-treatment” for this group are defined as number of hours        post Day 0 speculum exam.)

One Hour Post Treatment:

-   -   1. Vaginal pH (performed by research staff—two samples, two        readings)    -   2. Vaginal comfort assessment (subjects queried)    -   3. Adverse event reporting; targeted physical exam, if needed,        based on symptoms

Six Hours Post Treatment:

-   -   1. Vaginal pH (performed by research staff—two samples, two        readings)    -   2. Vaginal comfort assessment (subjects queried)    -   3. Adverse event reporting; targeted physical exam, if needed,        based on symptoms    -   4. Subject training to collect self-obtained vaginal swabs and        perform vaginal pH readings on Day 2 through Day 6.        (Verification of subject understanding and competency to        complete vaginal pH testing documented in source documents prior        to discharge of subject)

Twelve (+/−2) Hours Post Treatment:

-   -   1. Provide subject with diary to record vaginal pH readings,        vaginal comfort levels, and adverse events between visits    -   2. Vaginal pH (performed by subject, one sample, one reading)    -   3. Vaginal comfort assessment (subjects queried)    -   4. Adverse event reporting; targeted physical exam, if needed,        based on symptoms

Subjects who are not admitted on the same day as Screening, but within48 hours of Screening, have a urine pregnancy (UHCG) and vaginal pHtest, medical/surgical history, concomitant medications, AE assessmentand vital signs repeated upon admission administration of IP. If subjecthas consented to microbiome collection, the swab will again becollected.

Subjects admitted more than 48 hours but up to 5 days post Screeninghave a urine pregnancy (UHCG), vaginal pH test, speculum exam, vaginalsmear for Amsel criteria, medical/surgical history, concomitantmedications, AE assessment and vital signs repeated upon admittanceprior to administration of IP. If subject has consented to microbiomecollection, the swab will again be collected.

Day 1 (at the domiciliary unit): All subjects will remain in thedomiciliary unit for assessment of vaginal pH, vaginal comfort, andadverse reactions for a period of 24 (+/−2) hours, or longer asdetermined by the clinical trial site principal investigator. Thefollowing will be performed by the research staff and/or recorded in thesource documents and CRF:

-   -   1. Vaginal pH and microbiome collection (performed by research        staff) at 24 (+/−2) hours post-treatment    -   2. Demonstrate to subjects who have consented to microbiome        collection, the procedure of placing swab in transport        container, snapping off of the swab stick, closing lid securely.        Instruct subjects to place upright in their −20 freezer for        storage    -   3. Vaginal comfort assessment at 24 (+/−2) hours post-treatment    -   4. Review subject diary card at 24 (+/−2) hours post-treatment    -   5. Adverse event reporting at 24 (+/−2) hours post-treatment    -   6. Review concomitant medications at 24 (+/−2) hours        post-treatment    -   7. Vital signs measurement (BP, HR, temperature) at 24 (+/−2)        hours post-treatment    -   8. Laboratory Testing, performed by research staff at 24 (+/−2)        hours post-treatment (Vaginal pH, Wet Mount Smear, Whiff test,        all performed from vaginal swab—Amsel criteria evaluation from        results of these three tests) (Swab for vaginal pH reading is        collected prior to introduction of speculum for gynecologic        exam)    -   9. Gynecologic examination (speculum, performed by clinician) at        24 (+/−2) hours post-treatment    -   10. Targeted physical examination, if needed based on symptoms    -   11. Provide supplies to subject for self-obtained vaginal swabs        and vaginal pH readings for Days 2 through Day 6    -   12. Schedule Day 7 Final Study Visit    -   13. Subject released from domiciliary unit

Day 2 (Subject at home): The following will be performed and/or recordedin the source documents and CRF:

-   -   1. Adverse event reporting (subject records on diary card)    -   2. Vaginal comfort assessment (subject records on diary card)    -   3. Vaginal pH (performed by subject) at 48 (+/−4) hours        post-treatment (subject records on diary card).    -   4. In consented subjects, microbiome collection with storage of        swab in freezer

Day 3 (Subject at home): The following will be performed and/or recordedin the source documents and CRF:

-   -   1. Adverse event reporting (subject records on diary card)    -   2. Vaginal comfort assessment (subject records on diary card)    -   3. Vaginal pH (performed by subject) at 72 (+/−4) hours        post-treatment (subject records on diary card).    -   4. In consented subjects, microbiome collection with storage of        swab in freezer

Day 4 (Subject at home): The following will be performed and/or recordedin the source documents and CRF:

-   -   1. Adverse event reporting (subject records on diary card)    -   2. Vaginal comfort assessment (subject records on diary card)    -   3. Vaginal pH (performed by subject) at 96 (+/−4) hours        post-treatment (subject records on diary card).    -   4. In consented subjects, microbiome collection with storage of        swab in freezer

Day 5 (Subject at home): The following will be performed and/or recordedin the source documents and CRF:

-   -   1. Adverse event reporting (subject records on diary card)    -   2. Vaginal comfort assessment (subject records on diary card)    -   3. Vaginal pH (performed by subject) at 120 (+/−4) hours        post-treatment (subject records on diary card).    -   4. In consented subjects, microbiome collection with storage of        swab in freezer

Day 6 (subject at home): The following will be performed and/or recordedin the source documents and CRF:

-   -   1. Adverse event reporting (subject records on diary card)    -   2. Vaginal comfort assessment (subject records on diary card)    -   3. Vaginal pH (performed by subject) at 144 (+/−4) hours        post-treatment (subject records on diary card)    -   4. In consented subjects, microbiome collection with storage of        swab in freezer

Day 7 (+/−24 hours post treatment) [final study visit at the outpatientcenter of clinical trial site]: The following will be performed by theresearch staff and recorded in the source documents and CRF:

-   -   1. Review subject diary card    -   2. Adverse event reporting    -   3. Review concomitant medications    -   4. Query subject on current vaginal comfort    -   5. Laboratory Testing (Vaginal pH, Microbiome collection (in        consented subjects), Wet Mount Smear, Whiff test, all performed        from clinician obtained vaginal swab—Amsel criteria evaluation        from results of these three tests) (Swab for vaginal pH reading        is collected prior to introduction of speculum for gynecologic        exam)    -   6. Gynecologic exam (speculum, performed by clinician)    -   7. Targeted physical examination, if needed based on symptoms

In order to allow flexibility to subjects unable to attend the finalstudy visit on Day 7, a grace period of +/−24 hours is allowed for thisvisit when necessary (168+/−24 hours post treatment).

Unscheduled Visit(s): The subject will be asked to return to theclinical research site for evaluation if she develops symptoms or signsof illness and needs to be evaluated between scheduled visits. A medicalhistory will be taken and a targeted physical examination as dictated bythe symptoms. Safety lab or other lab tests will be obtained as deemednecessary by the Site PI. Findings will be documented in the sourcedocuments and on an eCRF for unscheduled visits. Adverse eventassessment (including SAEs) will be recorded in the source documents andon the eCRF.

Early Termination: For any subject who has received any amount of thestudy product and then withdraws or is withdrawn, an early terminationvisit will be conducted if the subject is willing. All activities listedfor Day 7 will be carried out unless medically contraindicated.

Clinical Laboratory Specimen Preparation, Handling and Storage: Alllaboratory specimens collected at JHU or MHMC during study visits willbe collected and processed by a trained study research team member,following site SOPs and all applicable site and local safety committeesafety guidelines. All samples collected on-site will be tested in theresearch unit; blood for HIV testing will be sent directly to the sitelaboratory for testing.

Safety Reporting and Safety Monitoring

Regulatory requirements including the FDA regulations, ICH Guidelinesfor Good Clinical Practice, and EU Clinical Trials Directive set forthsafety monitoring and reporting responsibilities of sponsors andInvestigators to ensure the safety and protection of human subjectsparticipating in clinical trials.

Responsibilities: The Site PIs participating in this clinical trial areresponsible for and will:

-   -   Evaluate subject safety including assessment of adverse events        (AEs) for seriousness, severity, and causality,    -   Notify ClinicalRM and the Sponsor, Evofem, Inc., of SAEs within        24 hours of awareness of event,    -   Provide detailed written reports, including necessary        documentation requested by the Sponsor or IRB/IEC, promptly        following immediate initial reports, and    -   Inform the IRB/IEC of AEs as required by applicable regulatory        requirements

Adverse Event (AE): Any untoward medical occurrence in a clinicalinvestigation subject who has received a study product intervention andthat does not necessarily have to have a causal relationship with thestudy product. An AE can, therefore, be any unfavorable and unintendedsign (including an abnormal laboratory finding, for example), symptom,or disease temporally associated with the use of a study medicinalproduct, whether or not considered related to the study medicinalproduct.

Serious Adverse Event (SAE): An SAE is any adverse event that results inany of the following outcomes:

-   -   Death;    -   Life-threatening (immediate risk of death);    -   In-patient hospitalization or prolongation of existing        hospitalization;    -   Persistent or significant disability or incapacity;    -   Congenital anomaly/birth defect;    -   Important medical events that may not result in death, be life        threatening, or require hospitalization may be considered a        serious adverse event when, based upon appropriate medical        judgment, they may jeopardize the subject and may require        medical or surgical intervention to prevent one of the outcomes        listed in this definition. Examples of such medical events        include allergic bronchospasm requiring intensive treatment in        an emergency room or at home, blood dyscrasias or convulsions        that do not result in in-patient hospitalization, or the        development of drug dependency or drug abuse.    -   The term “life-threatening” in the definition of SAE refers to        an event in which the subject was at risk of death at the time        of the event. It does not refer to an event which hypothetically        might have caused death if it were more severe.    -   The term “hospitalization” describes a period of at least 24        hours. Overnight stay for observation, treatment at an emergency        room or treatment on an out-patient-basis does not constitute a        hospitalization. However, medical judgment must always be        exercised and when in doubt, the case should be considered        serious.

Non-serious Adverse Event: A non-serious AE is any AE which does notfulfill the definition of an SAE.

Pre-treatment Adverse Event: A pre-treatment AE is any untoward medicaloccurrence arising or observed between informed consent and theinitiation of the study product.

Treatment Emergent Adverse Event (TEAE): A TEAE is any adverse eventoccurring after the application of the study product and within the timeof residual product effect, or a pre-treatment event or pre-existingmedical condition that worsens in intensity after the application ofstudy product and within the time of residual product effect. The timeof residual product effect is the estimated period of time after thedose of the study product, where the effect of the product is stillconsidered to be present based on change of vaginal pH from baseline.

Unexpected Adverse Reaction: An adverse reaction, the nature or severityof which is not consistent with the applicable product information(e.g., IND Safety Data).

Safety Reporting Requirements

Reporting Interval: All AEs and SAEs will be collected and recordedfollowing enrollment. The clinical trial site principal investigatorwill follow all AEs and SAEs until resolution (return to pretreatmentstatus or stabilization of conditions deemed as chronic) even if thisextends beyond the study-reporting period. At any time after thecompletion of the study, if a clinical trial site principal investigatorbecomes aware of a serious adverse event that is suspected to be relatedto the study product, the event will be reported to ClinicalRM, within24 hours of awareness. ClinicalRM will report the event immediately tothe Sponsor. The principal investigator will further ensure that theappropriate IRB has been notified of the reported SAE in a timely mannerper internal IRB reporting requirements.

Notification of the Sponsor of Serious Adverse Events: Any AE that meetsa protocol-defined serious criterion, must be submitted by sites within24 hours of site awareness on an SAE form to ClinicalRM, at thefollowing fax number and uploaded to the ClinicalRM Sponsor SharePointSite: Other supporting documentation of the event may be requested bythe Sponsor, Evofem, Inc. or ClinicalRM and should be provided as soonas possible. The medical monitor and clinical project manager will benotified of the SAE. The medical monitor will review and assess the SAEfor regulatory reporting and potential impact on study subject safetyand protocol conduct and inform the study Sponsor, Evofem, Inc.,according to the specified times for reporting AEs. Evofem, Inc. willinform the regulatory authorities and independent ethics committees(IECs/institutional review boards (IRBs)) in accordance with the localrequirements in force and International Conference on Harmonization(ICH) Good Clinical Practice (GCP) [ICH, GCP 2012]. ClinicalRM willnotify the Sponsor of any unexpected SAE with suspicion of being relatedto study drug in accordance with the local requirements. In addition,the Sponsor will be informed of any study related procedure SAE whichmay warrant a change of any study procedure or halting of the study.

Regulatory Reporting for Studies Conducted Under IND: Followingnotification from the Site PI, the Sponsor will report events that areboth serious and unexpected and that are associated with studyproduct(s) to the Food and Drug Administration (FDA) within the requiredtimelines as specified in 21 CFR 312.32: fatal and life-threateningevents within 7 calendar days (by phone or fax) and all other SAEs inwriting within 15 calendar days. All serious events designated as “notassociated” to study product(s) will be reported to the FDA at leastannually in a summary format.

Investigator's Assessment of Adverse Events: The determination ofseriousness, severity, and causality will be made by the Site PI who isqualified (licensed) to diagnose adverse event information, provide amedical evaluation of adverse events, and classify adverse events basedupon medical judgment. This includes but is not limited to physicians,physician assistants, and nurse practitioners.

Assessment of Seriousness: Event seriousness will be determinedaccording to the protocol definition of an SAE.

Assessment of Severity: Event severity will be assigned according to theTable of Clinical Events (AEs) for this study. The event may change inseverity before resolving. Highest severity per 24 hour block of timeand relevant changes in severity will be recorded.

Assessment of Adverse Event Causality: All AEs will be assessed by theSite PI for relationship to the trial product, independent of whetherthe adverse event is serious or non-serious AEs, and independent of itsseverity, using the following directions:

-   -   Definite: Good reasons and sufficient documentation to assume        direct involvement of the study drug in a causal relationship.    -   Probable: Good reasons and sufficient documentation to assume a        causal relationship.    -   Possible: A causal relationship is conceivable and cannot be        dismissed.    -   Unlikely: The event is most likely related to an etiology other        than the study product    -   Unrelated: The event is related to an etiology other than the        study product.

Adverse Events of Interest: In a double blinded, Phase 1 clinical safetytrial six subjects who were treated with the exemplary gel formulation(e.g., Acidform or Amphora® Gel) I reported no complaints and novulvovaginal or cervical irritation. No evidence of such inflammationwas observed by clinicians on visual or colposcopic inspection after sixdaily applications of IVAG the exemplary gel formulation (e.g., Acidformor Amphora® Gel) in a previous study. In a Phase I, placebo-controlled,randomized, closed-label study involving 14 days of twice-daily productexposure to the exemplary gel formulation (e.g., Acidform or Amphora®Gel) or KY® Jelly intravaginally twice-daily for 14 consecutive daysbetween menses. About two thirds of the exemplary gel formulation (e.g.,Acidform or Amphora® Gel) group compared to about one-third of the KY®Jelly group reported at least one symptom of genital irritation. Thesesymptoms, in general, were mild and included genital pain (burning,irritation, etc.), pruritus, pelvic cramping, vaginal candidiasis, andspotting. The three latter symptoms occurred later in the study (aftermultiple twice daily dosing). Safety data from a much larger Phase IIIstudy showed similar minor adverse events when the exemplary gelformulation (e.g., Acidform or Amphora® Gel) gel was applied regularly

Adverse Event's Outcome Measures: The outcome of all AEs will bereported based on the following definitions, independent of whether theyare serious or non-serious AEs, their severity, or their relationship tothe trial product:

-   -   Recovered: Fully recovered, or by medical or surgical treatment        the condition has returned to the level observed at the first        study related activity after the subject signed the informed        consent.    -   Recovering: The condition is improving and the subject is        expected to recover from the event. This term should only be        used when the subject has completed the study.    -   Recovered with sequelae: As a result of the AE, the subject        suffered persistent and significant disability/incapacity (e.g.        became blind, deaf, paralyzed). Any AE recovered with sequelae        should be rated as an SAE.    -   Not recovered    -   Fatal    -   Unknown: This term should only be used in cases where the        subject is lost to follow-up.

Process for Documentation of Events of Interest: Based on the safetyprofile of the exemplary gel formulation (e.g., Acidform or Amphora®Gel) observed in previous Phase I, Phase II, and Phase III studies, anassessment plan for anticipated adverse events will be implemented. AllAEs will be documented on the standard adverse event forms. Adverseevents forms for cutaneous AE grade 3 or higher will be emailed to theSponsor's representative, ClinicalRM, within 24 hours (non-serious orserious). The reporting of an AE grade 3 or higher will prompt follow-updiagnostic work-up (e.g., physical examinations, vital signs,descriptive assessment, subject diary and clinical laboratory work). Ifthe cutaneous AE is grade 3, subjects will be seen twice weekly untilthe severity is reduced to grade 2 or less at two consecutive visits andnot getting worse.

Other Safety Considerations: Any significant worsening or new findingsnoted during the final gynecological examination or any other potentialsafety assessments performed during the course of the study, whether ornot they are required by the protocol, should be recorded on theappropriate AE form.

Pregnancy: Woman of childbearing potential can be enrolled. However,women must agree to abstain from sexual intercourse over the duration ofthe study period. If there is any question that a subject will not bereliable in the adherence to this requirement, she should not be enteredinto the study. Study subjects must be instructed to notify the clinicaltrial site principal investigator immediately if they suspect pregnancyduring the course of the study and for a period of four weeks followingstudy discharge. The Site PI must report any pregnancy during the studyto ClinicalRM. Study subjects will give consent upon enrollment that theSite PI will report any pregnancy during the study to ClinicalRM andthat she will be asked to provide information about her pregnancy,delivery, and the health of her infant until age one month. The Site PImust report information on pregnancy and follow-up within 14 calendardays of obtaining the information using the Pregnancy Form and thePregnancy Follow-up Form, respectively. Pregnancy complications must berecorded as AEs. If the infant has a congenital anomaly or birth defect,the anomaly or defect must be reported and followed as an SAE.

Safety Oversight Contingency Plan: Due to the Sponsor's experience withthe IP and awareness of SAE probability based on past study data, noSafety Monitoring Committee (SMC) will be established for this study. Inthe event that SAEs occur at a rate different than anticipated, thestudy will be halted and a SMC initiated.

Follow-up of Adverse Events: During and following a subject'sparticipation in a clinical study, the Site PI/institution should ensurethat adequate medical care is provided to the subject for any AEs,including clinically significant laboratory values related to the study.The Site PI/institution should inform the subject when medical care isneeded for AEs of which the Site PI becomes aware. The post-treatmentfollow-up period for reporting new adverse events will continue untilthe last follow-up visit. All non-serious AEs classified as severe orpossibly/probably related or definitely related to the study productmust be followed until the subject has recovered and all queries havebeen resolved. However, cases of chronic conditions can be closed withan outcome of “recovering” or “not recovered”. If a subject dies fromanother event, these cases can be closed with an outcome of “notrecovered.” All other non-serious AEs must be followed until the outcomeof the event is “recovering” (for chronic conditions), or until the PIconsiders the AE to be stable, whichever comes first, and until allqueries related to these AEs have been resolved. If a subject dies fromanother event, these cases can be closed with an outcome of “notrecovered.” The clinical trial site principal investigator must ensurethat the worst case severity and seriousness is kept consistent throughthe series of AE forms and related AE follow-up form(s). The principalinvestigator must enter follow-up information about non-serious AE onthe AE form within the CRF. Queries or follow-up requests fromClinicalRM should be responded to within 14 calendar days, unlessotherwise specified. The Site PI must forward follow-up information onSAEs to ClinicalRM within 5 calendar days of obtaining the request forfollow-up information. All SAEs must be followed until the outcome ofthe event is recovered, recovered with sequelae or fatal and until allqueries have been resolved. For cases of chronic conditions and canceror if the subject dies from another event follow-up until the outcomecategories are “recovered”, “recovered with sequelae” or “fatal” is notrequired, as these cases can be closed with an outcome of “recovering”or “not recovered.”

Halting Criteria/Rules: The safety halting criteria rules are:

-   -   1. Death of an enrolled subject unless unequivocally not        attributable to study drug (e.g., traumatic injury)    -   2. Occurrence of a life-threatening allergic/hypersensitivity        reaction (anaphylaxis) within 48 hours of dosing, requiring        hemodynamic support with vasoactive medications or mechanical        ventilation the signs/symptoms will include any of the        following: bronchospasm, dyspnea, wheezing, stridor, hypoxemia        urticaria, angioedema, hives, and facial or oropharyngeal edema    -   3. An overall pattern of symptomatic or clinical events that the        sponsor, site PI or Medical Monitor consider associated with        study drug and that may appear minor in terms of individual        events, but that may collectively represent a serious potential        concern for safety.

If trial is prematurely terminated or suspended, the Sponsor shallinform investigators, regulatory authorities and IRB of the reason forhalting.

Example 2: A Randomized, Placebo Controlled Pilot Study to Determine theEffect and Duration of One Exemplary Gel Formulation (e.g., Acidform orAmphora® Gel) on Vaginal pH

Protocol of Example 1 was followed, unless otherwise indicated in thepresent example. The following amendments were made to the protocol ofExample 1:

-   -   The randomization process was amended from the methodology        stated in the protocol of Example 1. The protocol specified that        the biostatistician would transfer the randomization listing to        a study pharmacist. However, this process was modified to        specify that the biostatistician would transfer the        randomization listing to a designated unblinded data management        administrator prior to the start of the study.    -   To allow for flexibility to both subjects and domiciliary staff,        the following windows were permitted for sample collections at        post-treatment time points: 1 hour+30 minutes, 6 hours±30        minutes, 12 hours±2 hours, 24 hours±2 hours.    -   For sample collections at baseline and post-treatment at 1, 6,        12, and 24 hours, up to 3 swab collections could be taken to        allow for measurement of pH using pH papers covering different        pH ranges.    -   Analysis of the data on vaginal microbiome was handled by the        microbiome investigator.

One hundred and five volunteer women (at least 20 per treatment group)were treated with either Acidform, 5 g dose (Group A), Acidform, 4 gdose (Group B), Acidform, 3 g dose (Group C), placebo, 4 g (Group D), orno treatment (Group E). For the 3 treatment groups receiving Acidform(at either 3, 4, or 5 g), treatment was defined as speculum examinationplus administration of Investigational Product (IP). For the placebotreatment group, treatment was defined as speculum examination plusadministration of placebo. For Group E, the control, no treatment, wasdefined as speculum examination only.

Because vaginal pH values have been shown to vary by ethnicity, theintention was to recruit at least 13 subjects in each group who were ofeither African American or Hispanic descent. Routine screening testswere performed on admission, and subjects were assessed for asymptomaticBV via vaginal swabs obtained for grading by Amsel criteria.

A direct vaginal pH reading from each subject was obtained by researchstaff prior to the speculum examination, as well as 1 hour, and 6-hourspost-treatment (Day 0). Direct vaginal readings taken at the 1 hour and6-hours post-treatment time points were taken on specimens collectedfrom 2 different positions in the vagina, to allow for potentiallyincomplete distribution of the IP or placebo immediately followingadministration. Both readings were included in the full data analysissets. At the 6-hour post-treatment time point subjects were trained onself-collecting vaginal swabs and how to perform the vaginal pH test. At12-hours post-treatment, whilst still in the domiciliary clinic,subjects performed the vaginal pH test themselves using self-obtainedswabs, and recorded their results for clinician review.

Clinicians collected adverse event (AE) assessment data while subjectswere in the domiciliary unit, at each post-treatment vaginal pH testingtime point (1, 6, 12, and 24-hours post-treatment).

Additional vaginal microbiome collection was carried out for all thewomen who gave consent. The same swab for vaginal pH atBaseline/Screening, post 24-hour treatment and Days 2 through 7 wereused for the microbiome analysis. Research staff obtained the readingsfrom swabs taken at Baseline/Screening (a repeat pH/microbiome swab wastaken if screening and admission to the domiciliary unit did not occuron the same day), post 24-hour treatment and Day 7 (Table 4).

Subjects stayed overnight in the domiciliary units, and vaginal pH,microbiome collection (from consented subjects), and Amsel criteria weremeasured by research staff at 24-hours post-treatment prior to dischargeon Day 1. Subjects were discharged with the appropriate pH testingsupplies and a diary. All women had to agree to abstain from sexualintercourse, douching, and use of any intravaginally applied products ordevices until after their final study visit on Day 7.

Subjects measured their vaginal pH and carried out microbiome collectionat 24 (±4) hour intervals for 5 days (Days 2 to 6) as outpatients andrecorded the pH test results, any change in vaginal comfort, andconfirmation of having taken the microbiome swab in the diary provided.Subjects also recorded any activities engaged in, along with the daythis occurred, that were listed in the study exclusion criteria.

On Day 7 (±24 hours), subjects returned to the clinic with their diaryand swab transport containers. Clinic staff measured subjects' vaginalpH and Amsel criteria, and questioned them about any vaginal comfort(vaginal comfort assessment) over the course of the study (as per theself-recording in their diaries).

The Intervention and Follow-Up Period was defined as Day 0 to Day 7 inthe Schedule of Events (Table 4). The protocol-defined period ofobservation was 8 days (FIG. 2 ).

TABLE 4 Study Schedule Study Day Study Event −5 to 0 Screening visit 0Enrollment and admittance to domiciliary unit Administration of studytreatment 1 Inpatient follow-up and discharge 2-6 Subjectself-assessments at home 7 ± 1 Outpatient follow-up visit

TABLE 5 Study Protocol Chart Procedure Day 01- Day 01- Day 01- Day 01-Upon 1 hr Post 6 hr Post 12 hr Post Screening/Enrollment Admit AdmitAdmit Admit Day 1 Visit Type Screening Hospital Hospital HospitalHospital Hospital (clinic) admit admit admit admit admit Writteninformed X consent/HIPAA Evaluation X inclusion/exclusion criteriaDemographic data X Medical/surgical X X¹ history¹ Body height and Xweight¹ Concomitant X X¹ medication Vital signs (BP, X X¹ HR,temperature)¹ Gynecologic exam¹ X X¹ (speculum) Vaginal comfort X  X X XX assessment (subject recorded) Vaginal pH¹ X X   X^(2,6)  X^(2,6) X^(2,6) (research staff obtained) Vaginal X X  X microbiome (researchstaff obtained) Vaginal pH (self-  X⁶ obtained) Vaginal microbiome(subject obtained swab) Amsel criteria¹ X X¹ X (research staff obtainedswab) Urine pregnancy  X¹ X¹ test¹ HIV serology X Treatment (Groups X A, B, C, D, and E)³ Adverse event X  X X X X reporting (if applicable)Subject dairy X review Targeted physical X  X exam (if needed based onsymptoms) Procedure Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Visit Type N/AN/A N/A N/A N/A N/A home home home home home home Written informedconsent/HIPAA Evaluation inclusion/exclusion criteria Demographic dataMedical/surgical history¹ Body height and weight¹ Concomitant Xmedication Vital signs (BP, HR, temperature)¹ Gynecologic exam¹ X(speculum) Vaginal comfort X X X X X X assessment (subject recorded)Vaginal pH¹ X (research staff obtained) Vaginal X microbiome (researchstaff obtained) Vaginal pH (self-  X⁴  X⁴  X⁴  X⁴  X⁴ obtained) VaginalX X X X X microbiome (subject obtained swab) Amsel criteria¹ X (researchstaff obtained swab) Urine pregnancy test¹ HIV serology Treatment(Groups A, B, C, D, and E)³ Adverse event X X X X X X reporting (ifapplicable) Subject dairy X review Targeted physical X exam (if neededbased on symptoms) ¹Screening, Enrollment and admittance to domiciliaryunit for dosing (Day 0) could occur on the same day. Alternatively, oncescreened, subjects could be enrolled and admitted for dosing (Day 0) upto post screening Visit: Subjects who were not admitted on the same dayas Screening, but within 48 hours of Screening, had a urine pregnancy(UHCG) and vaginal pH test, medical/surgical history, concomitantmedication, and vital signs repeated upon admission. If the subjectconsented to Microbiome collection, the swab was again collected.Subjects admitted more than 48 hours up to 5 days post Screening had aUHCG, vaginal pH test, speculum exam, vaginal smear for Amsel criteria,medical surgical history, concomitant medication, and vital signs uponadmittance. If the subject consented to Microbiome collection, the swabwas again collected. ²For sample collections at baseline andpost-treatment at 1, 6, 12, and 24 hours, up to 3 swab collections maybe taken to allow for measurement of pH using pH papers coveringdifferent pH ranges. ³Groups A, B and C—IP (Acidform) administeredfollowing all screening tests and baseline vaginal pH test; GroupD—Placebo administered following all screening tests and baselinevaginal pH test; Group E—no treatment following all screening tests andvaginal pH test. ⁴In order to allow flexibility to subjects in theirdaily schedules, pH readings at X-hours post treatment ±4 hours isallowed. ⁵In order to allow flexibility to subjects unable to attend thefinal study visit on Day 7, a grace period of ±24 was allowed for whenthis visit was necessary. ⁶In order to allow flexibility for bothsubjects and domiciliary staff, the following windows were permitted forsample collection at post treatment: 1 hour ± 30 minutes, 6 hours ± 30minutes, 12 hours ± 2 hours, 24 hours ± 2 hours. BP = blood Pressure,HIPAA = Health Insurance Portability and Accountability Act, HIV = humanimmunodeficiency virus, Hosp = Hospital, HR = heart rate, N/A = notapplicable.

Study Design, Including the Choice of Control Groups

This was a Phase 1, randomized, placebo-controlled, double-blind,multi-center study.

Subjects were randomized in a 1:1:1:1:1 fashion across the 5 treatmentgroups. Simple randomization was performed using SAS®, and alldocumentation of this procedure and output was saved with the studybiostatistician's files until the end of the study. Prior to the startof the study, the randomization list was generated by the studybiostatistician and transferred to a designated unblinded datamanagement administrator. There was planned randomization for 15alternates, if required.

This study had 5 treatment groups: Acidform 5 g, Acidform 4 g, Acidform3 g, placebo 4 g, and no treatment. Study subjects in Groups A, B, C,and D (Acidform 5 g, 4 g, 3 g, and placebo, respectively) remainedblinded to their treatment assignment throughout the entire duration ofthe study. The Sponsor, Evofem, Inc., prepared IP as well as placebo,and supplied these products in ready to dispense status.

Three doses of Acidform (3 g, 4 g, and 5 g) were chosen, based onprevious studies, to assess if there was any difference in the effecteach of them had on vaginal pH and the duration of this effect.Universal placebo gel (UPG) was chosen as a comparator. In addition, ano-treatment group was included, not only to differentiate between theeffect the 3 doses of Acidform, placebo and no treatment had on vaginalpH, but also to monitor any natural changes in vaginal pH and microbiomeover a 7-day period (the duration of the study).

Study was double-blinded to remove any potential bias in the results.

This study was conducted at 2 sites across the United States (US) andresults were compiled separately for each center and combined foroverall results.

Selection of Study Population

Inclusion Criteria

To be considered eligible, all the following criteria must have been metat baseline:

1. Female subjects between 18 and 45 years, inclusive;

2. Ability to understand the consent process and procedures;

3. Agree to be available for all study visits;

4. Written informed consent in accordance with institutional guidelines;

5. Negative pregnancy test;

6. Able and willing to comply with all study procedures;

7. Have not engaged in sexual intercourse, douching, or used any form ofvaginal suppository or intravaginal device for 24 hours prior toenrollment;

8. Agree to abstain from sexual intercourse, douching, or any form ofvaginal suppository or intravaginal device use during the course of thestudy;

9. Report menstrual cycle regularity (25-day to 35-day menstrualcycles);

10. Test negative for BV or are positive but asymptomatic. (BV positivesubjects will be referred for treatment at or following the Day 7follow-up visit).

Exclusion Criteria

Subjects who met any of the following criteria at baseline wereineligible for participation in the study:

1. Participation in any study with an investigational compound or devicewithin 30 days prior to signing informed consent form;

2. Active drug or alcohol use or dependence that, in the opinion of theinvestigator, would interfere with adherence to study protocol;

3. Any other medical condition(s) that, in the judgment of theinvestigator, might interfere with the study or require treatment thatmight interfere with the study;

4. Family member of the investigation study staff;

5. Pregnant or breastfeeding;

6. Inability to provide informed consent;

7. A subject with a history or expectation of noncompliance withmedications or treatment protocol;

8. Women with symptoms of urinary tract infection (UTI), symptomatic BV,yeast infection or STI reported or observed during examination or basedon laboratory testing performed;*

9. Women who regularly use douches, vaginal medications orsuppositories, feminine sprays, genital wipes or contraceptivespermicides, or report abnormal vaginal discharge in the 48 hours priorto screening;

10. Women who are menstruating or who would expect to menstruate duringthe study;

11. Women who are currently using contraceptives that are directlydelivered to the vaginal mucosa, such as NuvaRing;

12. Any specific condition that, in the judgment of the investigator,precludes participation because it could affect subject safety.

*Should the study clinician make a presumptive diagnosis of suchinfections, these subjects will be withdrawn, referred for standard ofcare treatment, but will not be followed as part of this study. They maybe re-screened for potential enrollment 14 days after treatment perprotocol. Prohibited Medications, if all other screening criteria aremet.

Removal of Subjects from Therapy or Assessment

All subjects had the right to withdraw formal consent without prejudiceat any time during the study. If a subject withdrew formal consent, theinvestigator was to make a reasonable effort to determine the cause forwithdrawal of consent. For these subjects, as well as all other subjectswho required permanent discontinuation of study drug, the investigatorwas to make a reasonable effort to complete all required studyprocedures.

The subject's participation in this clinical study may have beenprematurely discontinued as a result of:

-   -   Development of any Exclusion Criteria;    -   Pregnancy or breastfeeding;    -   Request by subject to terminate participation;    -   Requirement for prohibited treatment (see Exclusion Criteria)        before Day 7;    -   Treatment-related toxicity;    -   Failure to adhere to requirements of the protocol including        treatment and safety monitoring;    -   Lost to follow-up;    -   Request of primary care provider;    -   At the request of the IRB/Ethics commit, FDA, or the Sponsor;    -   Incarceration;    -   The subject's well-being, based on the opinion of the Site PI.

If a subject discontinued study participation prematurely, the followingprocedures were to be completed:

-   -   Reason for discontinuation must be documented in the source        documents and eCRFs;    -   Subject to be contacted and encouraged to return for a follow-up        visit (with subject's consent) for safety;    -   Subject asked to complete an end-of-study evaluation (all        procedures from Final Study Visit, Day 7).    -   If an AE or SAE occurred, protocol-specified safety follow-up        procedures should be undertaken, and the subject encouraged to        receive appropriate care under medical supervision until the        symptoms of any AE resolve or the subject's condition becomes        stable.

Treatments

Treatments Administered

Study treatments were provided in pre-filled, single use applicators,sealed in a foil overwrap. The research study kit contained a group ofoverwrapped applicators placed into a standard white cardboard box alongwith instruction for use.

This was a single dose study, with study drug administered by the studyclinician on Day 0. Subjects were double-blinded to receive eitherAcidform 5 g, 4 g, or 3 g, or placebo 4 g. Subjects in the no-treatmentgroup received no treatment and were aware that they were not receivingIP.

Identity of Investigational Product(s)

The following products were used in this study and provided blinded bythe Sponsor:

Acidform; active ingredients: L-lactic acid, USP; citric acid, USP; andpotassium bitartrate, USP. All 3 active ingredients are designatedGenerally Regarded As Safe (GRAS) (21 CFR Part 184) and found in manyFDA-approved vaginally applied products in percentages similar to orless than those utilized in Acidform.

Acidform, 3 g, Lot number #00138, manufactured by Swiss AmericanProducts, Carollton, Tex.

Acidform, 4 g, Lot number #00139, manufactured by Swiss AmericanProducts, Carollton, Tex.

Acidform, 5 g, Lot number #00140, manufactured by Swiss AmericanProducts, Carollton, Tex.

Placebo; ingredients: Hydroxyethylcellulose (Natrasol 250 HX Pharm),sodium chloride, sodium hydroxide, sorbic acid and purified wateradjusted to pH 4.5; Lot number #KAC-C, manufactured by DPT Laboratories,San Antonio, Tex.

Method of Assigning Patients to Treatment Groups

The randomization was performed using SAS®, and all documentation ofthis procedure and output was saved with the study biostatistician'sfiles until the end of the study. The randomization list was generatedby the study biostatistician and transferred to a designated unblindeddata management administrator prior to the start of the study.

A total of 105 subjects were randomized in a 1:1:1:1:1 fashion acrossthe 5 study groups. Up to 15 alternates were available to replace studysubjects that dropped out.

Selection of Doses in the Study

The IP, Acidform, has been studied as an intravaginally administeredcontraceptive gel in the 5 g pre-filled applicator presentation (Studyof the Contraceptive Efficacy and Safety of Amphora® Gel Compared toConceptrol Vaginal Gel, Phase III, ClinicalTrials.gov ID #NCT01306331,completion date June 2014).

Three single doses of Acidform were chosen for this study, 5 g, 4 g, and3 g. This study assessed the effect of a single dose of Acidform onvaginal pH over 7 days. Several dose finding studies involving Acidformhave already been conducted for contraception.

A 4 g dose of standard placebo gel used in clinical trials was chosenfor use in this study as an inert comparator. It is an isotonicnon-buffering gel, pH adjusted to 4.5. In clinical trials, this placebohas been shown to be safe and acceptable when used up to twice daily for14 days.

Inclusion of a no-treatment group allowed for direct comparisons to seeif Acidform at any of the 3 doses (3 g, 4 g, or 5 g) had an effect onvaginal pH. In addition, comparison between any of the doses of Acidformwith no treatment enabled PIs to observe any potential effect ofAcidform on the prevention of BV in subjects without BV or who wereasymptomatic.

Selection and Timing of Dose for Each Patient

This was a single dose study. The study product (Acidform, 3 g, 4 g, 5g, or placebo) was administered intravaginally by the study clinician onDay 0, following subject admission to the domiciliary unit. Geladministration occurred using the pre-filled applicator after thegynecologic examination.

Blinding

Study subjects in Groups A, B, C, and D (Acidform 5 g, 4 g, 3 g, andplacebo, respectively) remained blinded to their treatment assignmentfor the entire duration of the study. The Sponsor, Evofem, Inc.,prepared IP as well as placebo, and supplied these products in ready todispense status.

The process for the preparation of the randomization for the studyensured that the study biostatistician was blinded to study treatmentassignments. The study biostatistician remained blinded until after thestudy database had been finalized and locked, and written instructionswere provided for unblinding.

Prior and Concomitant Therapy

Standard medical treatment(s) taken by the subject upon study entry weremaintained throughout the study. Concomitant medications were assessedand included any medication taken during the study, including all overthe counter medications, vitamins, and nutritional supplements.Information collected for each concomitant medication included, at aminimum: start date, stop date, or continuing, and indication.

Subjects must not have been currently taking or applying, either oral orintravaginal, or have taken or applied for 14 days prior to screening,any antibiotics, antimycotics, or probiotic compounds.

Treatment Compliance

Since the study drug was administered by the study clinician on Day 0,there was 100% treatment compliance across all groups.

Clinicians collected AE assessment data at each post-treatment vaginalpH testing time point while subjects were in the domiciliary unit (1, 6,12, and 24-hours post-treatment).

Subjects were discharged on Day 1 with the appropriate pH testingsupplies and a diary. On Day 2 through Day 6, inclusive, subjectscollected their own vaginal swabs and recorded their vaginal pH, and anyAE in the subject diary. On Day 7, subjects returned to the clinic fortheir Final Study Visit. Compliance was assessed using the diary andcollected vaginal swabs.

Efficacy and Safety Variables

Efficacy and Safety Measurements Assessed and Flow Chart

Summaries for the vaginal pH data were provided for each evaluation timepoint by treatment group and overall, as well as for all activetreatment groups combined. These summaries included the changes frombaseline.

Comparisons across the treatment groups with respect to change frombaseline in vaginal pH were performed using analyses of variance(ANOVA). The incorporation of the baseline vaginal pH score into theseanalyses was explored. Post-ANOVA pairwise comparisons of each studytreatment versus the placebo treatment, and versus no treatment was alsoassessed.

Graphic displays were compiled to illustrate the mean vaginal pH pertreatment group at each assessment time point, and the mean change frombaseline in mean vaginal pH per treatment group at each post-baselineassessment time point.

Amsel criteria were assessed as a potential indicator of BV infection.These assessments consisted of YES/NO indications for the presence ofeach of the following:

-   -   Vaginal pH>4.5;    -   Presence of thin, grayish white vaginal discharge;    -   Positive wet prep;    -   Positive whiff amine test.

Based on these individual criteria, a derived overall result(positive/negative) was reported. Categorical summaries for each Amselcriterion, as well as for the overall result, was compiled for eachassessment time point by treatment group and overall, as well as for allactive treatment groups combined.

Details of the assessments and procedures conducted throughout the studyare provided by procedure/assessment type and day, in Table 5.

Appropriateness of Measurements

The efficacy and safety measurements in this example are widely used andgenerally recognized as reliable, accurate, and relevant.

Primary Efficacy Variable(s)

Primary Efficacy Variables

The primary efficacy endpoint was assessment of change in vaginal pH andthe duration of this change from baseline, post-administration of asingle dose of Acidform or placebo, or no treatment.

Secondary Efficacy Variables

Secondary efficacy endpoints included the following:

-   -   Assessment of the effect of a single dose of Acidform or        placebo, or no treatment on asymptomatic BV, based on Amsel        criteria.    -   Assessment of the effect of a single dose of Acidform on the        vaginal microbiome.

Data Reporting

Data for this study included reported symptoms (including vaginalcomfort levels), AEs, clinical laboratory data, gynecologicalexamination, whiff test, wet prep specimen, and clinician andsubject-obtained pH values.

Clinical data was entered directly from the source documents, includingthe subject diary, to the data management system. CRAs performed sourceverification and query generation. CRAs worked with the Data Manager andsite personnel to freeze data prior to analysis. AEs, medical history,and medications were coded using MedDRA® 17 (or higher) and WHO drugdictionaries.

Study Patients

Disposition of Patients

In total 105 subjects were enrolled in the study and all subjects wererandomized 1:1:1:1:1 to one of the 5 treatment groups. All subjectsreceived treatment, either a single dose of IP or placebo, or notreatment on Day 0.

Five subjects discontinued the study, all from Acidform treatmentgroups: 3 subjects were lost to follow-up and 2 subjects discontinuedbecause menses came on during the study (categorized as “other” in Table6). No subjects discontinued the study due to an AE or SAE.

TABLE 6 Patient Disposition Acidform Acidform Acidform Placebo No- (5 g)(4 g) (3 g) (4 g) Treatment n = 22 n = 21 n = 21 n = 20 n = 21 Number ofsubjects Total Enrolled 22  21  21  20 21 Total Randomized 22  21  21 20 21 Total Treated 22  21  21  20 21 Included in Safety Population 22(100%)  21 (100%)  21 (100%)  20 (100%) 21 (100%) Completed 21 (95.5%)19 (90.5%) 19 (90.5%) 20 (100%) 21 (100%) Discontinued 1 (4.5%) 2 (9.5%)2 (9.5%) 0 0 Reasons for Discontinuation Adverse Event 0 0 0 0 0Protocol Violation 0 0 0 0 0 Withdrawal of Consent 0 0 0 0 0 Lost toFollow-Up  1 (100%)  2 (100%) 0 0 0 Other 0 0  2 (100%) 0 0 The SafetyPopulation included all subjects who received any amount of studytreatment, and all subjects who were randomized to the no-treatmentgroup. Percentages for primary categories are based on the number ofsubjects who were enrolled to the indicated treatment group. Percentagesfor subcategories are based on the number of subjects in the primarycategory being described. Percentages for reasons for discontinuationare based on the total number of subjects who discontinued among thosesubjects enrolled in the treatment group being summarized.

Demographic and Other Baseline Characteristics

A summary of demographic and baseline characteristics for the populationis presented in Table 7.

TABLE 7 Demographics and Baseline Characteristics for Overall StudyPopulation Acidform Acidform Acidform Placebo No- (5 g) (4 g) (3 g) (4g) Treatment n = 22 n = 21 n = 21 n = 20 n = 21 Age (years) n 22  21 21  20  21  Mean (SD) 31.62 (7.11) 28.57 (7.20) 28.12 (5.40) 30.42(6.47) 30.21 (6.45) Median   30.30   27.10   26.40   31.20   30.00 Min,Max (20.4, 45.6) (18.8, 44.5) (19.0, 38.9) (19.7, 45.0) (22.1, 46.0)Gender Male 0 0 0 0 0 Female 22 (100%) 21 (100%) 21 (100%) 20 (100%) 21(100%) Race White 10 (45.5%) 6 (28.6%) 11 (52.4%) 11 (55.0%) 7 (33.3%)Black or African American 10 (45.5%) 11 (52.4%) 8 (38.1%) 6 (30.0%) 12(57.1%) American Indian/Alaskan Native 0 0 0 0 0 Native Hawaiian orOther 0 0 0 0 0 Pacific Islander Asian 1 (4.5%) 0 1 (4.8%) 0 1 (4.8%)Other 1 (4.5%) 4 (19.0%) 1 (4.8%) 3 (15.0%) 1 (4.8%) Ethnicity Hispanicor Latino 6 (27.3%) 6 (28.6%) 2 (9.5%) 6 (30.0%) 3 (14.3%) Not Hispanicor Latino 16 (72.7%) 15 (71.4%) 19 (90.5%) 14 (70.0%) 18 (85.7%)Percentages are based on the number of subjects in the indicatedtreatment group.

Efficacy Results and Tabulations of Individual Subject Data

Efficacy results showing the effect and duration of a single dose of IP(Acidform at either 5 g, 4 g, or 3 g), placebo 4 g, and no treatment onvaginal pH from baseline through to Day 7 (final study visit) arepresented (FIG. 3 and FIG. 4 ).

Analysis of Efficacy

The mean vaginal pH for each treatment group and all assessment timepoints is shown in FIG. 3 .

For those subjects who received placebo, mean vaginal pH fluctuatedbetween 5.0 and 5.5, where subjects receiving no treatment had a meanvaginal pH around 4.8 to 4.7. In contrast, all subjects receivingAcidform at any dose had a noticeable decline in mean vaginal pH, within12 hours of the administration compared to the baseline pH measurementstaken at the time of administration. It was observed that the meanvaginal pH for subjects in any of the 3 Acidform treatment groupsremained lower than the mean vaginal pH of the no-treatment and theplacebo groups, upto Day 6 following administration. The greatestreductions in mean vaginal pH were seen with Acidform at the 5 g and 4 gdoses.

The mean change in vaginal pH from baseline through to Day 7, includingall intermediary time points, was recorded. These results formed part ofthe exploratory analysis to measure the duration of effect of a singledose of IP (Acidform at either 5 g, 4 g, or 3 g dose), placebo 4 g, andno treatment on vaginal pH (see FIG. 4 ).

For subjects who received the placebo or no treatment, no noticeablemean change in vaginal pH from baseline (measurements taken at the timeof administration) was observed. In contrast, all subjects receivingAcidform had a noticeable mean change in vaginal pH from the time ofadministration of Acidform to 12 hours compared to the baseline pHmeasurements taken at the time of administration. The mean change invaginal pH compared to baseline for subjects in any of the 3 Acidformtreatment groups remained noticeable compared to the mean change invaginal pH from baseline of the no treatment and the placebo group. Thegreatest reductions in mean vaginal pH were seen with Acidform at the 5g and 4 g doses. (FIG. 4 ).

Assessment of the effect of a single dose of Acidform or placebo, or notreatment on asymptomatic BV based on Amsel criteria, was assessed as anexploratory endpoint. Results of these findings were assessed on subjectadmittance, Day 1 and Day 7. Although 4 of 22 (19%) of subjects in theAcidform 5 g treatment group had positive Amsel findings at baseline,none had positive Amsel findings at Day 1. By Day 7, 1 of 22 (5%) hadpositive Amsel findings. This finding suggests Acidform 5 g preventsrecurrence of BV infection, at least in the short term.

Adjustments for Covariates

The comparison across study treatments with respect to changes frombaseline vaginal pH values explored the use of baseline pH as acovariate. Other than this possible application, no adjustments wereplanned for this study.

Handling of Dropouts or Missing Data

No imputation of missing data values was performed. All analyses werebased on data as observed.

Efficacy Results

A single dose of Acidform at any of the 3 studied doses lowered vaginalpH from baseline, and this mean reduction was significantly greater thanthat seen with placebo or no treatment. Peak reduction occurred at12-hours post-administration, with the greatest reduction in meanvaginal pH from baseline seen with the highest (5 g) dose. Subjects inthe Acidform treatment groups continued to have a lower mean vaginal pHat Day 6 compared to the mean vaginal pH of subjects in the placebo andthe no-treatment group. Subjects in the Acidform treatment groupscontinued to have mean reduction in vaginal pH at Day 7 compared tobaseline.

Safety Evaluation

A global summary of demographics of the safety population is presentedin Table 8. A detailed summary of AEs, including subjects with at leastone treatment-emergent adverse event (TEAE), treatment-related AEs,SAEs, and AEs leading to study withdrawal during the 7-day doubleblindtreatment period of the study is presented in Table 9. The incidence ofAEs by body system organ class and preferred term is summarized in Table8 for the treatment period. There were no SAEs reported in this studyand no AEs led to study withdrawal during the treatment period. AEs arereported using MedDRA® (Version 17.0 or higher).

Extent of Exposure

A total of 105 subjects participated in this study and were randomizedacross 5 treatment groups. Of the 105 subjects, only 84 receivedtreatment (including placebo) as 21 subjects were assigned to theno-treatment group. As this was a single dose study, all subjectsenrolled were included in the safety population (Table 8).

Adverse Events

Brief Summary of Adverse Events

Sixty-eight subjects (64.8%) reported a TEAE, none of which wereserious, severe or lifethreatening, and did not lead to studydiscontinuation. TEAEs ranged from 61.9% to 76.2% across the treatmentgroups including IP or placebo, and was 47.6% in the no-treatment group.

Three subjects (14.3%) in the no-treatment group were assessed by studyclinicians as having a treatment-related TEAE, which is explained by thefact that this was a double-blind study and could imply that some TEAEsexperienced by subjects are naturally experienced by females over time.

TABLE 8 Overview of Treatment-Emergent Adverse Events (SafetyPopulation) Acidform Acidform Acidform Placebo No- (5 g) (4 g) (3 g) (4g) Treatment Category n = 22 n = 21 n = 21 n = 20 n = 21 TEAEs 16(72.7%) 13 (61.9%) 16 (76.2%) 13 (65.0%) 10 (47.6%) Treatment-RelatedTEAEs¹ 12 (54.5%)  8 (38.1%) 11 (52.4%)  7 (35.0%)  3 (14.3%) Severe orLife-Threatening TEAEs 0 0 0 0 0 Serious TEAEs 0 0 0 0 0 Serious,Treatment-Related TEAEs 0 0 0 0 0 TEAEs Leading to Study Treatment 0 0 00 0 Modification or Discontinuation from Study TEAE = Treatment-EmergentAdverse Event ¹Included TEAEs were considered by the Investigator to beprobably or definitely related to the study drug. Subjects may have morethan one AE per category. For each category, a subject was counted onceif she reported one or more events. Percentages are based on the numberof subjects in the indicated treatment group. The Safety Populationincluded all subjects who received any amount of study treatment, andall subjects who were randomized to the no-treatment group.

TABLE 9 Overview of Treatment-Emergent Adverse Events by System OrganClass (Safety Population) Acidform Acidform Acidform Placebo No- SystemOrgan Class (5 g) (4 g) (3 g) (4 g) Treatment Preferred Term n = 2.2 n =21 n = 21 n = 20 n = 21 Total Number of AEs 34  28  32  26  13  TotalNumber of Subjects with at Least One AE 16 (72.7%) 13 (61.9%) 16 (76.2%)13 (65.0%) 10 (47.6%) Infections and Infestations 1 (4.5%) 0 0 0 1(4.8%) Nasopharyngitis 0 0 0 0 1 (4.8%) Urinary Tract Infection 1 (4.5%)0 0 0 0 Musculoskeletal and connective tissue disorders 1 (4.5%) 0 0 0 0Muscle spasms 1 (4.5%) 0 0 0 0 Nervous system disorders 2 (9.1%) 0 0 1(5.0%) 1 (4.8%) Headache 1 (4.5%) 0 0 1 (5.0%) 1 (4.8%) Sciatica 1(4.5%) 0 0 0 0 Total Number of AEs 34  28  32  26  13  Renal and urinarydisorders 0 1 (4.8%) 1 (4.8%) 0 0 Micturition urgency 0 1 (4.8%) 0 0 0Pollakiuria 0 0 1 (4.8%) 0 0 Reproductive system and breast disorders 16(72.7%) 13 (61.9%) 16 (76.2%) 13 (65.0%) 9 (42.9%) Cervical discharge 1(4.5%) 1 (4.8%) 2 (9.5%) 2 (10.0%) 3 (14.3%) Cervix disorder 1 (4.5%) 01 (4.8%) 2 (10.0%) 1 (4.8%) Cervix inflammation 0 0 1 (4.8%) 0 0Dysmenorrhea 0 2 (9.5%) 0 0 0 Menstruation irregular 1 (4.5%) 0 1 (4.8%)0 0 Vaginal discharge 15 (68.2%) 13 (61.9%) 12 (57.1%) 10 (50.0%) 3(14.3%) Vaginal hemorrhage 0 1 (4.8%) 1 (4.8%) 3 (15.0%) 1 (4.8%)Vaginal odor 0 0 2 (9.5%) 0 0 Vulvovaginal burning sensation 1 (4.5%) 01 (4.8%) 0 0 Vulvovaginal discomfort 1 (4.5%) 0 2 (9.5%) 0 0Vulvovaginal pain 1 (4.5%) 1 (4.8%) 0 1 (5.0%) 2 (9.5%) Vulvovaginalpruritis 3 (13.6%) 2 (9.5%) 1 (4.8%) 3 (15.0%) 0 The total number of AEscounts all AEs for all subjects. Subjects may have more than one AE perbody system and preferred term. At each level of subject summarization,a subject was counted once if she reported one or more events.Percentages are based on the number of subjects in the indicatedtreatment group. AE = adverse event.

Display of Adverse Events

The most common AE across all treatment groups was vaginal discharge(Table 9). This finding was noted in the majority of subjects whoreceived IP (Acidform at any dose or placebo), ranging from 50.0% to68.2%, and 14.3% of subjects in the no-treatment group (Table 9).Gel-associated “discharge” peaked on Day 2 and declined thereafter, andis best explained by leakage of some of the gel over time.

No other frequent AEs were noted or were of any cause for concern.

Adverse Events by Severity

All AEs reported in this study were mild to moderate, across alltreatment groups: the majority were classified as mild.

Relationship of Adverse Event to Study Drug

AE rates were similar across the four treatment groups, but were lowerfor the no-treatment group. There appeared to be no significantdifference in AE frequency or severity between the 3 doses of Acidform(5 g, 4 g, and 3 g), or between the 3 doses of Acidform and placebo.

Severity of Adverse Events Related to Study Drug

All related AEs reported in this study were mild to moderate, across alltreatment groups.

Deaths, Other Serious Adverse Events, and Other Significant AdverseEvents

There were no deaths, SAEs, or other significant AEs reported in thisstudy.

Safety Conclusions

Sixty-eight subjects (64.8%) reported a TEAE, none of which wereserious, severe or life-threatening. The most common AE across alltreatment groups was vaginal discharge, which was higher among subjectsreceiving IP (Acidform at either 5 g, 4 g, or 3 g, or placebo) comparedwith subjects receiving no treatment. Administration of gel, bothplacebo and Acidform, resulted in leakage of some gel over time that wasexperienced as “discharge”, which diminished over time.

No deaths or SAEs were reported in this study. All reported TEAEs weremild or moderate and did not require concomitant treatment or lead tostudy discontinuation.

Discussion and Overall Study Conclusions

Vaginal pH was lowered from baseline with a single dose of Acidform atthe 3 studied dosages (5 g, 4 g, and 3 g). The mean reduction wassignificantly greater than reductions in vaginal pH seen with eitherplacebo or no treatment. Peak reduction occurred 12-hourspost-administration of Acidform, with the greatest reduction in meanvaginal pH from baseline seen with the highest (5 g) dose. Subjects inthe Acidform treatment groups continued to have mean reduction invaginal pH at Day 7 compared to baseline.

Certain Embodiments

Embodiment 1 provides a method of treating bacterial vaginosis (BV)comprising intravaginally administering a composition to a subject withBV, wherein the composition comprises: (a) a polymer thickener; (b)L-lactic acid; and (c) a preservative, wherein administering thecomposition ameliorates symptoms of BV in the subject.

Embodiment 2 provides a method of reducing the recurrence rate ofbacterial vaginosis (BV) comprising intravaginally administering acomposition to a subject with BV, wherein the composition comprises: (a)a polymer thickener; (b) L-lactic acid; and (c) a preservative, whereinadministering the composition reduces recurrence rate of bacterialvaginosis in the subject.

Embodiment 3 provides a method of preventing the recurrence of bacterialvaginosis (BV) comprising intravaginally administering a composition toa subject with BV, wherein the composition comprises: (a) a polymerthickener; (b) L-lactic acid; and (c) a preservative, whereinadministering the composition prevents recurrence of bacterial vaginosisin the subject.

Embodiment 4 provides the method of any one of embodiments 1-3, whereinthe preservative is selected from a group consisting of benzoic acid,sodium benzoate, methylparaben, ethylparaben, butyulparaben,propylparaben, benzyalkonium chloride, phenylmercuric nitate, andchlorhexidine.

Embodiment 5 provides the method of any one of embodiments 1-4, whereinthe polymer thickener is selected from a group consisting of xanthangum, alginic acid, hydroxypropyl cellulose, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, chitosan, polycarbophil, andcarbopol.

Embodiment 6 provides the method of any one of embodiments 1-5, whereinthe polymer thickener is a combination of xanthan gum and alginic acid.

Embodiment 7 provides the method of any one of embodiments 1-6, whereinthe composition further comprises a humectant.

Embodiment 8 provides the method of any one of embodiments 1-7, whereinthe humectant is selected from a group consisting of glycerol,polyethylene glycol, propylene glycol, sorbitol, and tiracetin.

Embodiment 9 provides the method of any one of embodiments 1-8, whereinthe composition further comprises a pharmaceutically acceptable carrier.

Embodiment 10 provides the method of any one of embodiments 1-9, whereinthe composition further comprises water.

Embodiment 11 provides the method of any one of embodiments 1-10,wherein the composition further comprises a buffer.

Embodiment 12 provides the method of any one of embodiments 1-11,wherein the buffer comprises citric acid and potassium bitartrate.

Embodiment 13 provides the method of any one of embodiments 1-12,wherein the composition is administered once during a course oftreatment.

Embodiment 14 provides the method of any one of embodiments 1-12,wherein the composition is administered multiple times during a courseof treatment.

Embodiment 15 provides the method of embodiment 14, wherein the courseof treatment comprises an initial course of treatment and a subsequentcourse of treatment and wherein the composition is administered morefrequently during the initial course of treatment than during thesubsequent course of treatment.

Embodiment 16 provides the method of embodiment 15, wherein the initialcourse of treatment is between 1 week and 4 weeks.

Embodiment 17 provides the method of embodiment 16, wherein the initialcourse of treatment is about 2 weeks to about 3 weeks.

Embodiment 18 provides the method of embodiment 17, wherein the initialcourse of treatment is about 1 week.

Embodiment 19 provides the method of any one of embodiments 15-18,wherein the subsequent course of treatment is about 1 week to about 19weeks.

Embodiment 20 provides the method of any one of embodiments 15-19,wherein the subsequent course of treatment is about 2 weeks to about 18weeks.

Embodiment 21 provides the method of any one of embodiments 15-20,wherein the subsequent course of treatment is about 3 weeks to about 17weeks.

Embodiment 22 provides the method of any one of embodiments 15-21,wherein the subsequent course of treatment is about 4 weeks to about 16weeks.

Embodiment 23 provides the method of any one embodiments 15-22, whereinthe subsequent course of treatment is 5 weeks to about 15 weeks.

Embodiment 24 provides the method of any one of embodiments 15-23,wherein the subsequent course of treatment is about 6 weeks to about 14weeks.

Embodiment 25 provides the method of any one of embodiments 15-24,wherein the subsequent course of treatment is about 7 weeks to about 13weeks.

Embodiment 26 provides the method of any one of embodiments 15-25,wherein the subsequent course of treatment is about 8 weeks to about 12weeks.

Embodiment 27 provides the method of any one of embodiments 15-26,wherein the subsequent course of treatment is about 9 weeks to about 10weeks.

Embodiment 28 provides the method of any one of embodiments 15-27,wherein the subsequent course of treatment is about 10 weeks to about 11weeks.

Embodiment 29 provides the method of any one of embodiments 15-28,wherein the frequency of administration during the initial course oftreatment is daily.

Embodiment 30 provides the method of any one of embodiments 15-29,wherein the frequency of administration during the initial course oftreatment is every other day.

Embodiment 31 provides the method of any one of embodiments 15-30,wherein the frequency of administration during the initial course oftreatment is once a week.

Embodiment 32 provides the method of any one of embodiments 15-31,wherein the frequency of administration during the initial course oftreatment is once every 2 weeks.

Embodiment 33 provides the method of any one of embodiments 15-32,wherein the frequency of administration during the subsequent course oftreatment is every other day.

Embodiment 34 provides the method of any one of embodiments 15-33,wherein the frequency of administration during the initial course oftreatment is once every week.

Embodiment 35 provides the method of any one of embodiments 15-34,wherein the frequency of administration during the initial course oftreatment is once every 2 weeks.

Embodiment 36 provides the method of any one of embodiments 15-35,wherein the frequency of administration during the initial course oftreatment is once every 3 weeks.

Embodiment 37 provides the method of any one of embodiments 1-36,wherein the composition is administered at a dosage from about 0.5 g toabout 10 g.

Embodiment 38 provides the method of any one of embodiments 1-37,wherein the composition is administered at a dosage from about 3 g toabout 5 g.

Embodiment 39 provides the method of any one of embodiments 1-38,wherein the composition is administered at a dosage selected from about3 g, about 4 g, and about 5 g.

Embodiment 40 provides a method of prognosis for risk of recurrence ofbacterial vaginosis (BV) in a subject with BV, the method comprising (a)testing vaginal pH of the subject; (b) intravaginally administering acomposition as defined in any one of embodiments 1-12 to the subject,(c) re-testing vaginal pH of the subject; and (d) prognosing risk ofrecurrence of BV based on comparing the results of the vaginal pH testsin step (a) and (c).

Embodiment 41 provides a method of prognosis for risk of recurrence ofbacterial vaginosis (BV) in a subject with BV, the method comprising (a)analyzing vaginal microbiome of the subject; (b) intravaginallyadministering a composition as defined in any one of embodiments 1-12 tothe subject, (c) re-analyzing vaginal microbiome of the subject; and (d)prognosing risk of recurrence of BV based on comparing the results ofthe vaginal microbiome analyses in step (a) and (c).

Embodiment 42 provides a method of prognosis for risk of recurrence ofbacterial vaginosis (BV) in a subject, the method comprising (a)assessing BV in a subject using Amsel criteria; (b) intravaginallyadministering a composition as defined in any one of embodiments 1-12 tothe subject, (c) re-assessing BV in the subject using Amsel criteria;and (d) prognosing risk of recurrence of BV based on comparing theassessments in step (a) and (c).

Embodiment 43 provides the method of any one of embodiments 40-42,wherein the composition is administered once during a course oftreatment.

Embodiment 44 provides the method of any one of embodiments 40-42,wherein the composition is administered multiple times during a courseof treatment.

Embodiment 45 provides the method of embodiment 44, wherein the courseof treatment comprises an initial course of treatment and a subsequentcourse of treatment and wherein the composition is administered morefrequently during the initial course of treatment than during thesubsequent course of treatment.

Embodiment 46 provides the method of embodiment 45, wherein the initialcourse of treatment is between 1 week and 4 weeks.

Embodiment 47 provides the method of embodiment 45, wherein the initialcourse of treatment is about 2 weeks to about 3 weeks.

Embodiment 48 provides the method of embodiment 47, wherein the initialcourse of treatment is about 1 week.

Embodiment 49 provides the method of any one of embodiments 45-48,wherein the subsequent course of treatment is about 1 week to about 19weeks.

Embodiment 50 provides the method of any one of embodiments 45-49,wherein the subsequent course of treatment is about 2 weeks to about 18weeks.

Embodiment 51 provides the method of any one of embodiments 45-50,wherein the subsequent course of treatment is about 3 weeks to about 17weeks.

Embodiment 52 provides the method of any one of embodiments 45-51,wherein the subsequent course of treatment is about 4 weeks to about 16weeks.

Embodiment 53 provides the method of any one embodiments 45-52, whereinthe subsequent course of treatment is 5 weeks to about 15 weeks.

Embodiment 54 provides the method of any one of embodiments 45-53,wherein the subsequent course of treatment is about 6 weeks to about 14weeks.

Embodiment 55 provides the method of any one of embodiments 45-54,wherein the subsequent course of treatment is about 7 weeks to about 13weeks.

Embodiment 56 provides the method of any one of embodiments 45-55,wherein the subsequent course of treatment is about 8 weeks to about 12weeks.

Embodiment 57 provides the method of any one of embodiments 45-56,wherein the subsequent course of treatment is about 9 weeks to about 10weeks.

Embodiment 58 provides the method of any one of embodiments 45-57,wherein the subsequent course of treatment is about 10 weeks to about 11weeks.

Embodiment 59 provides the method of any one of embodiments 45-58,wherein the frequency of administration during the initial course oftreatment is daily.

Embodiment 60 provides the method of any one of embodiments 45-59,wherein the frequency of administration during the initial course oftreatment is every other day.

Embodiment 61 provides the method of any one of embodiments 45-60,wherein the frequency of administration during the initial course oftreatment is once a week.

Embodiment 62 provides the method of any one of embodiments 45-61,wherein the frequency of administration during the initial course oftreatment is once every 2 weeks.

Embodiment 63 provides the method of any one of embodiments 45-62,wherein the frequency of administration during the subsequent course oftreatment is every other day.

Embodiment 64 provides the method of any one of embodiments 45-63,wherein the frequency of administration during the initial course oftreatment is once every week.

Embodiment 65 provides the method of any one of embodiments 45-64,wherein the frequency of administration during the initial course oftreatment is once every 2 weeks.

Embodiment 66 provides the method of any one of embodiments 45-65,wherein the frequency of administration during the initial course oftreatment is once every 3 weeks.

Embodiment 67 provides the method of any one of embodiments 40-66,wherein the composition is administered at a dosage from about 0.5 g toabout 10 g.

Embodiment 68 provides the method of any one of embodiments 40-67,wherein the composition is administered at a dosage from about 3 g toabout 5 g.

Embodiment 69 provides the method of any one of embodiments 40-68,wherein the composition is administered at a dosage selected from about3 g, about 4 g, and about 5 g.

Embodiment 70 provides the method of any one of embodiments 40 and43-69, wherein lowering of vaginal pH between steps (a) and (c) isassociated with good prognosis for reduced risk of recurrence of BV.

Embodiment 71 provides the method of any one of embodiments 41 and43-69, wherein restoration of a healthy vaginal microbiome between steps(a) and (c) is associated with good prognosis for reduced risk ofrecurrence of BV.

Embodiment 72 provides the method of any one of embodiments 42-69,wherein the assessment result is Amsel-positive or Amsel-negative in thesteps (a) and (c).

Embodiment 73 provides the method of embodiments 72, wherein change inassessment result from Amsel-positive to Amsel-negative between steps(a) and (c) is associated with good prognosis for reduced risk ofrecurrence of BV.

1. A method of treating bacterial vaginosis (BV) comprisingintravaginally administering a composition to a subject with BV, whereinthe composition comprises: (a) a polymer thickener; (b) about 2.5% w/vL-lactic acid; (c) a preservative, d) citric acid, (e) potassiumbitartrate, (f) a humectant, (g) a pH adjusting agent, and (h) asolvent, wherein the composition is administered multiple times during acourse of treatment, wherein the course of treatment comprises about 1week to about 20 weeks, wherein the composition is administered at adosage from about 0.5 g to about 10 g per dose, and whereinadministering the composition ameliorates symptoms of BV in the subject.2. The method of claim 1, wherein the solvent is water.
 3. The method ofclaim 1, wherein the course of treatment comprises an initial course oftreatment and a subsequent course of treatment and wherein thecomposition is administered more frequently during the initial course oftreatment than during the subsequent course of treatment.
 4. The methodof claim 3, wherein the frequency of administration during the initialcourse of treatment is daily, less than once daily, every other day,once a week, or once every 2 weeks.
 5. The method of claim 3, whereinthe frequency of administration during the subsequent course oftreatment is less than once daily, every other day, once a week, onceevery 2 weeks, or once every 3 weeks.
 6. The method of claim 1, whereinthe composition is administered at a dosage from about 3 g to about 5 g,about 3 g, about 4 g, or about 5 g.
 7. The method of claim 1, whereinthe subject has previously been diagnosed as Amsel-positive.
 8. Themethod of claim 7, wherein upon of administration of the composition thesubject is Amsel-negative, thereby treating the BV.
 9. A method ofreducing the recurrence rate of bacterial vaginosis (BV) comprisingintravaginally administering a composition to a subject, wherein thecomposition comprises: (a) a polymer thickener; (b) about 2.5% w/vL-lactic acid; (c) a preservative, (d) citric acid, (e) potassiumbitartrate, (f) of a humectant, (g) a pH adjusting agent, and (h) asolvent, wherein the composition is administered multiple times during acourse of treatment, wherein the course of treatment comprises about 1week to about 20 weeks, wherein the composition is administered at adosage from about 0.5 g to about 10 g per dose, and whereinadministering the composition reduces recurrence rate of bacterialvaginosis in the subject.
 10. The method of claim 9, wherein the subjectis a subject with BV.
 11. The method of claim 10, wherein the subjectwith BV is asymptomatic.
 12. The method of claim 9, wherein the subjectis free of BV.
 13. The method of claim 9, wherein the polymer thickeneris selected from a group consisting of xanthan gum, alginic acid,hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodiumcarboxymethyl cellulose, chitosan, polycarbophil, and carbopol.
 14. Themethod of claim 9, wherein the solvent is water.
 15. The method of claim9, wherein the course of treatment comprises an initial course oftreatment and a subsequent course of treatment and wherein thecomposition is administered more frequently during the initial course oftreatment than during the subsequent course of treatment.
 16. The methodof claim 15, wherein the frequency of administration during the initialcourse of treatment is daily, less than once daily, every other day,once a week, or once every 2 weeks.
 17. The method of claim 16, whereinthe frequency of administration during the subsequent course oftreatment is less than once daily, every other day, once a week, onceevery 2 weeks, or once every 3 weeks.
 18. The method of claim 9, whereinthe composition is administered at a dosage from about 3 g to about 5 g,about 3 g, about 4 g, or about 5 g.
 19. The method of claim 1, wherein apH less than 5 is maintained in the vagina.
 20. The method of claim 9,wherein a pH less than 5 is maintained in the vagina.